Age-related cognitive decline is a significant public health concern. Recently, non-pharmacological methods, such as physical activity and mental training practices, have emerged as promising low-cost methods to slow the progression of age-related memory decline. In this study, we investigated if Tai Chi Chuan (TCC) and Baduanjin modulated the fractional amplitude of low-frequency fluctuations (fALFF) in different frequency bands  and improved memory function. Older adults were recruited for the randomized study. Participants in the TCC and Baduanjin groups received 12 weeks of training (1 h/day for 5 days/week). Participants in the control group received basic health education. Each subject participated in memory tests and fMRI scans at the beginning and end of the experiment. We found that compared to the control group: (1) TCC and Baduanjin groups demonstrated significant improvements in memory function; (2) TCC increased fALFF in the dorsolateral prefrontal cortex (DLPFC) in the slow-5 and low-frequency bands; and (3) Baduanjin increased fALFF in the medial PFC in the slow-5 and low-frequency bands. This increase was positively associated with memory function improvement in the slow-5 and low-frequency bands across the TCC and Baduanjin groups. Our results suggest that TCC and Baduanjin may work through different brain mechanisms to prevent memory decline due to aging. Age is the main risk factor for most common neurodegenerative diseases, such as mild cognitive impairment (MCI) and Alzheimer’s disease (AD). Memory dysfunction is the primary cognitive symptom in MCI and AD and has a profound impact on those whom it affects  and Baduanjin are popular mind-body practices  between 0.01 Hz and 0.08 Hz are of particular relevance to resting state fMRI  50–70 years old; (2) right-handed; and (3) no regular physical exercise for at least 1 year . Exclusion criteria were: (1) history of stroke; (2) suffered from severe cerebrovascular disease, musculoskeletal system disease, or other contraindications caused by sports injury; (3) a score of Beck depression inventory (BDI-II) ≥ 14  was used to assess the memory function of each participant. The WMS-CR is designed to assess memory function  T1-weighted images were collected.All MRI scans were acquired on a 3.0T magnetic resonance scanner  with an 8-channel phased-array head coil. For the rs-fMRI, the scans were acquired with TR = 2100 ms, TE = 30 ms, flip angle = 90°, slice thickness = 3 mm, gap = 0.6 mm, acquisition matrix = 64 × 64, voxel size = 3.125 × 3.125 × 3.6 mmPost hoc analysis (Sidak corrected) was applied to explore the between-group differences.Baseline characteristics were compared by one-way analysis of variance (ANOVA) and Chi square tests using SPSS 18.0 Software . During the analysis, all control participants from the two cohorts were combined into one group to increase the power. In order to estimate the effects of TCC and Baduanjin, ANCOVA analysis was applied to compare the change of MQ and the subtests across the three groups with age (years), with gender and education (years) included as covariates in the model. http://rfmri.org/DPARSF; Chao-Gan and Yu-Feng, The fMRI data preprocessing was performed using Data Processing Assistant for Resting-State fMRI (DPARSF) Software  and the second factor had two levels (pre- and post-treatment). Age, gender and years of education were also included in the analysis as covariates of non-interest. A threshold of a voxel-wise 102 older adults between 50–70 years old were screened for this study. Of the 90 participants who were qualified for the study and finished baseline scans, 62 participants completed all study procedures . Four participants in the TCC group dropped out . Nine participants in the Baduanjin group dropped out (8 due to scheduling conflicts and 1 due to unwillingness to participate in the MRI scan). Fifteen participants in the control group dropped out (11 due to scheduling conflicts and 4 due to inability to participate in post-treatment MRI scans). One subject in the Baduanjin group was excluded from fALFF analysis due to excessive head movement (exceeded 3.0 mm).P > 0.05). Average attendance rates were 95% in the TCC group (ranging from 88% to 100%) and 97% in the Baduanjin group (ranging from 92% to 100%).Group characteristics are shown in Table F = 25.45, p < 0.001). Post hoc Sidak correction analysis showed that compared with the control group, MQ scores significantly increased in the TCC and Baduanjin groups . There were no significant differences between the TCC and Baduanjin groups (p = 0.233). The comparisons of the subscores of WMS-CR showed TCC significantly increased visual reproduction subscores compared to controls. Baduanjin produced greater improvement in mental control, recognition, visual reproduction, touch and comprehension memory subscores compared to controls after bonferroni correction (p < 0.0063). Baduanjin also produced greater improvements in touch subscores compared to TCC after bonferroni correction (p < 0.0063). Please also see our previous publications on subscore changes across different treatment  among the three groups showed that after 12 weeks, fALFF was significantly increased in the right DLPFC in the TCC group compared to the control group Table . In the p < 0.005 uncorrected with 10 continuous voxels. We found that compared to the Baduanjin group, there was a significant increase in fALFF in the periaqueductal gray, bilateral DLPFC, and temporoparietal junction in the TCC group. Compared with the TCC group, the Baduanjin group was associated with a significant fALFF increase in the left mPFC and left precuneus.To explore the difference between Tai Chi vs. Baduanjin intervention, we also applied a relatively less conservative threshold of voxel-wise r = 0.48, p = 0.005 significant after Bonferroni correction (0.025 (0.05/2); Figure p = 0.048, not significant after Bonferroni correction across the TCC and Baduanjin groups).To explore the association between the fALFF changes observed above and behavioral outcomes, we also extracted the average fALFF values of the significant clusters (DLPFC and mPFC) and performed a multiple regression analysis including age, gender, and education as covariates. Results showed a significant association between the fALFF changes at mPFC and corresponding MQ .To explore the association between the fALFF changes observed above and behavioral outcomes, we also extracted the average fALFF values of the significant clusters and performed multiple regression analysis respectively including age, gender and education as covariates across the participants in TCC and Baduanjin group. Results showed a significant association between the mPFC fALFF changes and corresponding MQ changes  were observed. When we applied a relatively less conservative threshold of voxel-wise In this study, we investigated the effects of 12 weeks of TCC and Baduanjin exercise on fALFF changes and clinical outcome measures in older adults. We found that: (1) MQ significantly increased in both TCC and Baduanjin groups compared with the control group; (2) TCC increased fALFF in the right DLPFC in the slow-5 band and the 0.01–0.08 Hz band; and (3) Baduanjin increased fALFF in the bilateral mPFC in the slow-5 band and the 0.01–0.08 Hz band following exercise. fALFF changes at the mPFC in the slow-5 and 0.01–0.08 Hz bands showed a significant positive association with corresponding MQ changes.Both TCC and Baduanjin are mind-body exercises consisting of meditation, breathing, and gentle movements. From the viewpoint of physical exercise, both TCC and Baduanjin are safe aerobic activities  Tai Chi vs. Control; and (2) Baduanjin vs. Control. Furthermore, we also found that TCC and Baduanjin modulate the subtests of WMS-CR differently, Baduanjin can significantly increase the touch subscore compared to TCC, which further suggests that different mechanism may underlying Tai Chi Quan and Baduanjin. We speculate this difference may due to different exercise characteristics associated with the two mind-body interactions. TCC involves more complicated body movements and requires moving the trunk and all four limbs, whereas the movement involved in Baduanjin is much simpler.We found that compared to controls, participants in the TCC group had increased fALFF in the right DLPFC, while the participants in the Baduanjin group had increased fALFF in the bilateral mPFC in the slow-5 band and the 0.01–0.08 Hz band. Although TCC and Baduanjin are associated with different patterns compared to controls, we did not find significant differences between the TCC and Baduanjin groups at this threshold we set. However, at a less conservative threshold of voxel-wise In this study, we found that TCC exercise increased fALFF in the right DLPFC in the slow-5 band and the 0.01–0.08 Hz band compared to the control group. Previous studies have demonstrated that the DLPFC, a task positive region, is a key area in the cognitive control network  showed fALFF changes after TCC and Baiduanjin practice, while no significant differences were observed in the slow-4 band. This suggests that the memory-relevant changes induced by 3 months of TCC and Baduanjin practice are specifically reflected difference low-frequency band. In a previous study, Han et al.  investigThe current study has several limitations. First, the sample size is relatively small. Second, both TCC and Baduanjin are considered mind-body exercises. Therefore, our design could not disentangle the effect of physical activity vs. mental exercise, and was unable to identify the crucial components of the exercise affecting memory improvement and brain functional fluctuation changes. Further research is needed to compare the effects of exercise, meditation, TCC and Baduanjin directly. Third, we did not record the participants’ original physical activity habits, and only gave them an introduction to keep their original physical activity habits throughout the duration of the experiment. Further study should record the participants original physical activity habits for more accurate documentation of their activity intensity.In this study, we found that 12 weeks of intensive group TCC/Baduanjin practice can significantly modulate fALFF in different frequency bands and improve memory function. TCC increased fALFF in the slow-5 band and the 0.01–0.08 Hz band in the DLPFC, and Baduanjin increased fALFF in the slow-5 band and the 0.01–0.08 Hz band in the mPFC. fALFF changes in the mPFC in the slow-5 band and 0.01–0.08 Hz band were also correlated with general memory improvement. Our results imply that TCC and Baduanjin may work through different brain mechanisms due to differences in the characteristics and complexity of their respective regimens, but both exercises may hold the potential to prevent age-related memory decline.The Medical Ethics Committee in the Affiliated Rehabilitation Hospital of Fujian University of Traditional Chinese Medicine approved all study procedures. The experiment was performed in accordance with approved guidelines. All participants signed a written consent. This study was registered on the Chinese Clinical Trial Registry (ChiCTR-IPR-15006131).LC: experimental design; JK: analysis and manuscript preparation; JT: experimental design, data analysis and manuscript preparation; GZ: data analysis; JL and XX: data collection and data analysis; WL, JW, ML, ZW and KH: data collection; XC and NE: manuscript preparation. All authors contributed to manuscript draft and have read and approved the final manuscript.JK has a disclosure to report; holding equity in the startup company MNT, but declares no conflict of interest. The other 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 FAM83 proteins were recently identified as novel transforming oncogenes that function as intermediaries in EGFR/RAS signaling. Using two distinct forward genetics screens, the Bissell and Jackson laboratories uncovered the importance of the FAM83 proteins in promoting resistance to EGFR tyrosine kinase inhibitors and therapies targeting downstream EGFR signaling effectors. The discovery of this novel oncogene family using distinct genetic screens provides compelling evidence that the FAM83 proteins are key oncogenic players in cancer-associated signaling when they are overexpressed or dysregulated. Consistent with a role in oncogenic transformation, the FAM83 genes are frequently overexpressed in diverse human cancer specimens. Importantly, ablation of numerous FAM83 members results in a marked suppression of cancer-associated signaling and loss of tumorigenic potential. Here, we review the current knowledge of the FAM83 proteins’ involvement in cancer signaling and discuss the potential mechanisms by which they contribute to tumorigenesis. Both redundant activities shared by all 8 FAM83 members and non-redundant activities unique to each member are highlighted. We discuss the promise and challenges of the FAM83 proteins as novel points of attack for future cancer therapies. Cell signaling networks are complex, interconnected pathways that work in unison to control cellular function. When these tightly controlled pathways are disrupted, however, cells may become cancerous –3. IdentHere, we discuss a novel family of signaling proteins, named FAM83 (FAMily with sequence similarity 83), that function as key intermediates in oncogenic EGFR, MAPK and PI3K/AKT signaling –25. Due One of the most promising potential roles for the FAM83 proteins is within the ErbB signaling network, both at the receptor level and the downstream MAPK and PI3K/AKT pathways. The ErbB signaling network tightly controls normal cell growth and proliferation . Often, The ErbB receptors and MAPK and PI3K/AKT signaling cascades are the subject of intense research aimed at identifying pharmacological inhibitors that will suppress growth signaling and prevent cancer cell proliferation. Precision therapies aimed at disrupting ErbB RTKs  , RAS Fa. In thiset al. used nested PCR to show that FAM83A mRNA is highly expressed in the circulating tumor cells (CTCs) of lung adenocarcinoma patients [et al. also identified FAM83A as a potential biomarker in breast cancer [The smallest FAM83 protein, FAM83A , was first implicated as a potential cancer biomarker in 2008. Liu patients . When ust cancer . Using aet al. utilized an elegant phenotypic reversion assay in 3D laminin-rich gel. In their screen, malignant HMECs, which grow as depolarized, disorganized, proliferative colonies in laminin-rich basement membrane (lrBM), can be reverted to a non-malignant phenotype  by EGFR TKIs [In addition to the potential use of FAM83A as a biomarker, a forward genetic screen identified FAM83A as a driver of EGFR tyrosine kinase inhibitor (TKI) resistance, suggesting FAM83A plays an important role in cancer cell biology . EGFR isGFR TKIs   receptor by binding to domain IV in the extracellular region. The drug functions through several mechanisms that include blockade of downstream signaling and prevention of HER2 cleavage into a catalytically active cytoplasmic fragment . Althoug problem . Boyer eve cells . This five cells . Thus, iSimultaneous to the discovery of FAM83A as a driver of TKI resistance, the Jackson laboratory discovered FAM83B as a driver of human mammary epithelial cell (HMEC) transformation . Using aFAM83B interacts with a noteworthy list of signaling proteins that facilitate its role as a signaling oncoprotein. For example, the Jackson laboratory has shown that FAM83B blocks the inhibitory interaction of 14-3-3 proteins with CRAF, promoting CRAF membrane localization and thereby inducing activation of MAPK signaling pathway and supporting tumor growth . The Jacin vivo tumorigenicity [The involvement of FAM83B in multiple signaling pathways strengthens its potential for precision therapeutic targeting. As mentioned, precision medicines have targeted the PI3K/AKT/mTOR and MEK/ERK signaling pathways. However, selective inhibition of MEK/ERK signaling has consistently induced compensatory activation of PI3K/AKT signaling and vice versa, ultimately conferring resistance. The sensible option would be to simultaneously target MAPK and PI3K/AKT/mTOR signaling, hoping to dampen the compensatory signaling. This approach continues to suffer from the added complexity of harming normal tissues that rely on these pathways. Given that FAM83B overexpression can induce multiple EGFR signaling pathways including EGFR/PLD, MAPK, PI3K/AKT/mTOR, we hypothesized that inhibition of FAM83B might suppress multiple pathways and prevent the compensatory survival signaling. Indeed, ablation of FAM83B in breast and colon cancer cells significantly suppressed tumor cell growth and genicity . The grogenicity .Not only does FAM83B activate MAPK and PI3K/AKT/mTOR signaling, but it may also participate in cancer-associated Wnt signaling. In a proteomics effort to identify important players in the Wnt signaling pathway, FAM83B was identified as a novel interacting partner of negative regulatory proteins Axin-1 and APC . While set al. recently linked elevated expression of FAM83D and its ability to promote MEK/ERK signaling with higher incidence of hepatocellular carcinomas (HCC) [Gene expression studies reveal that FAM83 members are not only elevated in a wide variety of cancers, but in many instances more than one of the FAM83 proteins are significantly elevated. For example, the Jackson laboratory recently described that FAM83A, B, D and E are all highly expressed in ovarian cancers, and similarly bladder cancers show elevated levels of FAM83A, C and D . While tas (HCC) , 71.via regulation of EGF-driven Src/AKT signaling pathways [In addition to redundant transformative properties, FAM83 proteins likely also have non-redundant functions. These proteins lack any significant homology beyond the DUF1669 and vary greatly in size ranging from 434-1179 amino acids  binds to the chromokinesin KID and localizes to the spindle during mitosis to regulate spindle maintenance and cell division . Accordiin silico, bioinformatics analysis on the C-terminal sequences in combination with cellular assays using C-terminal swap/deletion mutants of FAM83 proteins.Similar to the previously identified role of FAM83A and FAM83B in resistance to EGFR TKI inhibitors, newer reports implicate FAM83H in therapeutic resistance. An insertional mutagenesis screen to find genes that confer Erlotinib resistance in pancreatic cancers identified a common integration site, LOC100128338, which shares chromosomal location with FAM83H . TherefoWhile several FAM83 proteins have been implicated in receptor-mediated signaling pathways  and in modulating their downstream effectors, the mechanism by which elevated FAM83 expression hyperactivates each of these pathways remains unclear. Moreover, given the potential redundant and non-redundant functions described in the previous section, the manner in which each FAM83 protein contributes to oncogenic signaling may be unique. Here, we discuss potential modes of action for the FAM83 proteins.Evidence to date supports an adaptor/scaffold function for the FAM83 proteins since they lack obvious enzymatic functions, yet interact with essential signaling intermediaries. Current known FAM83 interacting proteins include EGFR, RAF, PI3K-p85, PI3K-p110, AKT, Axin-1, and APC , 68, 69.In addition to kinase signaling, FAM83 proteins may have additional adaptor functions in cancer cells, including nucleating and inactivating enzymes that destabilize oncogenic signaling proteins . FAM83D Analysis of the human proteome across most major tissue types indicates that FAM83 members are expressed at low (often undetectable) levels throughout the body relative to other proto-oncogenes such as EGFR and RAS Figure . When coA greater understanding of the mechanisms by which the FAM83 proteins enhance and prolong oncogenic signaling will also be critical for determining whether and how each FAM83 protein can be targeted. Studies have shown FAM83 proteins interact with many key cancer-signaling proteins including EGFR (FAM83B), PI3K (FAM83A-B), AKT (FAM83B), c-RAF (FAM83A-E) –25, 68. in silico attempts at disrupting adaptor interactions depend on sufficient knowledge of the protein structure and protein-binding interfaces, which are currently lacking for most FAM83 proteins. The only crystal structure solved is of FAM83A, which is the smallest FAM83 protein, largely consisting of the DUF1669 domain [Thus, while feasible, rational drug design and 9 domain . Again, 9 domain , 68. Forin vivo, with little immunogenicity, no changes in body weight, and no issues with clotting [Another method for targeting adaptor proteins is to suppress their expression using RNA interference (RNAi). RNA interference (RNAi) as a therapeutic regimen is continuously improving and holds great potential –108. siRclotting . The rapThe significant discovery of a new family of oncogenes as critical mediators of EGFR/RAS signaling provides the basis for the development of novel therapeutics that target FAM83 members in a wide variety of cancers. The tumor-exclusive increase in expression of FAM83 proteins makes them highly desirable targets. It is also clear that sensitivity to select targeted therapies is closely linked to the expression of the FAM83 oncogenes, and screening for FAM83 expression will help predict response to treatment strategies as well as to determine future therapeutic options. We envision future studies will involve identification of the unique function of each FAM83 protein, and development of new strategies to therapeutically target both the conserved and unique oncogenic properties of FAM83 proteins.


The role that serum uric acid (UA) plays in the pathophysiological development of erectile dysfunction (ED) is controversial. We aimed to screen the factors related with ED, and to examine the association between serum UA and ED. Our data were derived from a cross-sectional Survey on Prevalence in East China for Metabolic Diseases and Risk Factors study in 2014–2015. Questionnaire of International Index of Erectile Dysfunction-5 was used for assessment of ED. Data were collected in three general communities respectively. A total of 1365 men were enrolled with an overall mean age 55.5 ± 10.8 years (range: 20–83 years). The prevalence of ED was 62.4% (51.4% standardized) in the population. Males with ED were older, and more prone to have a higher follicle-stimulating hormone, luteinizing hormone, sex hormone-binding globulin, glycated hemoglobin, fasting plasma glucose levels and lower free androgen index (FAI), UA levels, and more likely to have diabetes and elevated blood pressure compared with those without ED. Age and UA were independent influencing factors for ED. Besides, UA was positively correlated with FAI after adjustment for age. In conclusion, our study demonstrated the protective role that UA might play in development of ED. Even though ED is not a fatal condition, it interferes with daily routine, social interactions and quality of life of the patient. ED is a common clinical problem, especially in aging men2, and the number of patients suffered from ED will be up to 322 million by 2025 worldwide3. Given the etiology of ED is multifactorial, pathophysiology of ED is influenced by several physical systems4.Erectile dysfunction (ED), a kind of male sexual dysfunction, is characterized by the inability to achieve or maintain erections of the penis potent for satisfaction during sexual activity5. So far, several experimental and clinical studies on the fields of the relationship between UA and ED have given clues on the potential causative role of UA in ED occurrence7. However, most of the above deductions were based on the researches which were restricted to patients with particular diseases, such as hypertension8 and coronary artery disease9, hence, the generalization of these studies are limited. To confirm the connection between ED and UA, large-scale and population-based studies which provide convincing evidence are needed.Uric acid (UA), the end product of dietary and endogenous purine metabolism, is currently confirmed to modulate the physiological functions of various physical systems. Several epidemiological cohort studies have demonstrated that the level of serum UA is associated with many clinical events, such as metabolic syndrome, hypertension, and diabetesA large population-based study, referred to as Survey on Prevalence in East China for Metabolic Disease and Risk Factors (SPECT-China), was launched in 2015–2016. Based on the analysis of the data from the survey, we attempted to explore the independent influential factors for ED and the linkage of ED with UA.Table th national population census data, www.stats.gov.cn). The percentiles of ED were initially low (38.2%) at ages less than 40 years and then increased progressively with increasing age. We noticed that the prevalence of ED increased an average 10 percent per decade approximately. On the contrary, the serum UA concentration showed a declining trend with the increase of age generally.Figure Table To explore the independent risk or protective factor for erectile function, binary logistic regression analysis was performed. Univariate logistic regression analysis showed that age  ; FSH1.054,1.031–1.077); LH ; SHBG ; HbA1c ; FPG ; systolic pressure  were significantly associated with increased odds of ED, on the contrary, both FAI  and UA  had protective effect on erectile function , and the OR of UA to ED was 0.998 (0.996–1.000). The results confirmed that age was an independent risk factor for ED, and UA was an independent protective factor  is similar to several population-based studies7. Additionally, the data of two studies focused on the males with hypertension8 and coronary artery disease9 respectively, revealed the detrimental role that UA played in the process of ED. The conclusions of the above studies are hard to be persuasive because several great limitations exist, for example, the results are not universal due to the small sample size and the research methods such as case-control are liable to cause selection bias. Their results may be applicable for specific population groups with particular disease, and the viewpoint that UA is a risk factor for ED lacks evidences derived from large population-based cross-sectional studies for supporting. By contrast, our demonstration of the association between UA and ED based on demographic analysis is more universal and representative. Although the mechanism is not clearly illustrated, we attempt to elucidate the linkage between UA and ED from the following points.Frankly speaking, one contentious area is whether UA plays a protective role for ED or not. Various reports holding the opposite view have described that UA was a potential risk factor for the presence of ED. In a case-control study, compared with the normal group, the serum UA levels of males with ED elevated significantly (P < 0.001)16. Meanwhile, recent studies have found low UA level was associated with depression18. In a case-control study, lower UA level was a characteristic of depression compared with the healthy group, in addition, the UA level raised to 312.28 μmol/L from the original 271.97 μmol/L after a five-week treatment with antidepressants (P = 0.043). Similarly, a multivariate analysis conducted by Gu et al. revealed that the declining serum UA promoted the development of depression19. Given the linkage between UA and depression, it provides an explanation on the protective role that UA plays in maintaining the erection of penis by alleviating depression.Initially, depression, a kind of psychogenic problems, is considered to be directly correlated with ED20. Oxidative stress impairs the vascular endothelium by enhancing production of reactive oxygen species (ROS)21, consequently contributes to the pathogenesis of ED22. Hyperglycemia induces abnormalities of vascular endothelium by promoting release of ROS24. Consistently, in our study, the men suffered from diabetes were with a high prevalence of ED. Besides, numerous studies have provided evidence that UA is a powerful scavenger of ROS25. For example, a randomized, double-blind, placebo-controlled study has reported that administration of UA raises circulating antioxidant defense and accelerates the restoration of endothelium-dependent vasodilation which represents the improvement of endothelial function26. Furthermore, a crossover study conducted by Waring has validated that substantial reduction of serum UA concentrations is invalid for restoring endothelial function in patients with diabetes27. Therefore, UA may be beneficial for erectile function through its powerful antioxidative effect on endothelium protection to some extent. And the viewpoint may give a reasonable explanation for the results in our study that the prevalence of ED decreases with the increasing concentration of UA. Moverover, with regard to the patients with diabetes, our study demonstrates that ED is more prevalent in the lower UA group than in higher UA group, which is consistent with Waring’s study.As a vascular event, penile erection requires an intact endothelium to occur, and endothelial dysfunction is a manifestation of ED28. In our study, we also confirmed that ED prevalence was inversely correlated with FAI, but not TT. An explanation is that the level of androgenicity may not be exactly reflected by TT due to the inactive part which binds with circulating SHBG29. Subsequently, we delineated the positive correlation between UA and FAI, which is in accordance with previous researches31. In Kurahashi’s study, after the androgen replacement therapy, the UA concentration increased and hyperuricemia was more prevalent in the high-androgen-dose group31. Trials on laboratory animals have demonstrated that androgens promote UA reabsorption in the kidney30. To sum up, androgens may prevent the deterioration of erectile function by raising the serum UA concentration.Androgens are indispensable in maintaining erectile response, and the underlying molecular mechanisms have been clarified clearly during the past decadeBased on our results of multivariate logistic regression analysis, serum UA is an independent protective factor for ED. The OR of UA is 0.998, which is very close to 1, indicating that the effect of UA on ED is modest. And this could partly explain why a very small numerical difference in UA concentrations exists between ED and non-ED groups (339.8 ± 77.4 vs 349.9 ± 76.2 μmol/L). In addition, we conclude that serum UA levels increase with age which supporting the major effect of age on UA. The mean age of men with ED group is distinctly higher than that of men without ED group. Hence, if age is adjusted, UA concentration in ED group would be lower than 339.8 μmol/L, and the difference of serum UA between the two groups would be larger.Our study had several strengths. First, quality of the study was controlled strictly, and the same trained research group completed all the anthropometric measurements and questionnaires, meanwhile, all the biochemical and hormonal items were examined in one laboratory certified by the College of American Pathologists. Second, our results are more universal because the study was performed with large sample size. However, some important limitations must be recognized. First, due to the cross-sectional nature of the study, we cannot identify the causality but only the association between serum UA level and ED. Second, this study recruited mainly Han Chinese men and the results cannot apply to other ethnic groups. Third, the sample in our study was a relatively older population with the mean age 55.5 years, and the sample size in each age group had a certain difference. Although International Index of Erectile Dysfunction-5 (IIEF-5) is widely recognized for assessment of ED in both clinical and epidemiologic studies, dividing sharply whether suffering from ED or not according to the IIEF-5 score below 22 is not appropriate. Lastly, ED, which is closely related with self-esteem, is a highly private topic for males. Hence, the possibility exists that parts of the participants didn’t fill in the IIEF-5 questionnaire honestly, and the uncontrolled factor may cause bias.In conclusion, based on this population-based cross-sectional study, we estimated the prevalence of ED in Chinese males. Age, SHBG, FAI, FSH, LH, HbA1c, FPG, diabetes, systolic pressure and UA had strong association with ED. Among them, UA was an independent protective factor of ED. In addition, UA had a notable positive correlation with FAI.www.chictr.org.cn)33. To obtain more persuasive and generalized data, a stratified cluster sampling method was adopted. Our investigation covered both rural and urban areas, meanwhile, different economic development of area was considered. This investigation was conducted in Shanghai, Anhui Province and Jiangsu Province. Followings were the inclusion criteria: male adults aged 18 years and older; living in the current area for 6 months or longer; and Chinese citizens. Participants were excluded if they unwilling to participate; received diagnosis of serious psychiatric diseases; used drugs that affected sexual function and experienced no sex life for 6 months prior to the date of the investigation.The survey, SPECT-China, is a population-based cross-sectional study on the prevalence and influential factors of metabolic disease. The registration number is ChiCTR-ECS-14005052 (Altogether 2178 participants took part in the study. The respondents who refused completing questionnaire (n = 414), had no sex life within the last six months (n = 287) and incomplete laboratory or questionnaire data (n = 112) were excluded. Finally, a total of 1365 men were enrolled in the study. Prior to the data collection of this research, all the participants signed written informed consent. The investigation was approved by the Ethics Committee of Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine. In accordance with the Declaration of Helsinki, the research was carried out.34. Subsequently, the collection of anthropometric data, such as body weight, height and blood pressure, was used standard methods as described previously35. To protect participants’ privacy and to ensure data authenticity and reliability, IIEF-5 questionnaires were completed in a separate room. To obtain the accurate biochemical indexes and hormone levels of blood samples, all the participants were asked to fast overnight at least 8 h before the study. The blood samples for the plasma glucose test were collected into anticoagulant vacuum tubes and centrifuged on the spot within 1 hour after collection. Blood samples were stored at −20 °C after being collected and shipped by air on dry ice. A central laboratory, certified by the College of American Pathologists, was responsible for the measurement of blood samples indexes. HbA1c was assessed by HPLC . FPG was measured by a Beckman CoulterAU680. Sex-related hormones including TT, LH and FSH were measured via chemiluminescence using the Immulite 2000 platform . The missing values of TT under the minimal detectable limit were replaced by the mean values that were 0.35 nmol/L between 0 and the minimal detectable limit. SHBG levels were detected via electrochemiluminescence using a Cobas E601 analyzer . BMI was calculated as weight in kilograms divided by height in meters squared. FAI, which may reflect increased androgenicity properly36, is the ratio of total circulating testosterone to SHBG.A standard procedure was executed by the same trained staff group to complete questionnaire. The questionnaire was split into two parts: one included information on demographic characteristics, medical history and lifestyle, the other is the IIEF-5 questionnaire, which is used to estimate the severity of ED34. Diabetes was defined as fasting plasma glucose ≥7.0 mmol/L, or HbA1c ≥6.5% based on the American Diabetes Association 2014 criteria.In regard of the assessment of ED, participants with the IIEF-5 questionnaire score below 21 were classified into the ED group, and the rest with score 22–25 were considered with normal erectile functionThe analysis of the collected data was performed with IBM SPSS Statistics, version 22 . The mean values and a number with proportion were adopted to summarize the general characteristics for continuous variables and categorical variables respectively. Kolmogorov–Smirnov analysis was used for normality testing. For abnormal distribution data, Mann–Whitney U test was used. A logistic regression analysis was used to evaluate the risk factors for ED. All considered variables were introduced in the univariate and multivariate model, meanwhile, OR and 95% CI were presented. Linear regression analysis was used for examining the association between UA and sex-related hormones. The conventional P < 0.05 was used to assess statistical significance.Supplementary Table S1


Cranial cruciate ligament (CCL) rupture is a common cause of pelvic limb lameness in dogs. The tibial plateau leveling osteotomy (TPLO) is a well-described surgical procedure that treats CCL ruptures. The objective of this study was to compare the risk of tibial tuberosity fractures from TPLO procedures using a TPLO reduction pin in situ versus patients with a TPLO reduction pin removed at the time of surgery. Our hypothesis is that patients with a TPLO reduction pin left in situ will have a decreased incidence of tibial tuberosity fractures.A total of 400 dogs that fitted the criteria of 200 consecutive TPLO surgeries performed with each group were included in the study. The Student’s t-test revealed a statistically significant difference in fractures observed in group 1 (in situ pin) and group 2 (no pin). In univariate logistic regression analysis, only the covariate for the presence of the reduction pin was associated with a statistically significant reduction in the likelihood of tibial tuberosity fracture. In the multivariate model, the presence of the reduction pin was associated with an approximate 92% reduction in the likelihood of tibial tuberosity fracture.The online version of this article (10.1186/s13104-018-3474-7) contains supplementary material, which is available to authorized users. Cranial cruciate ligament (CCL) rupture is one of the most common causes of pelvic limb lameness in dogs . The exaAvulsion fractures of the tibial tuberosity have been reported to occur in 1–9% of patients following a TPLO and may contribute to an increased morbidity and the need for revision surgery . It has Several surgeons from this institution have elected at times to leave the temporary TPLO reduction pin in situ following completion of the TPLO plate application. Although the placement of the temporary anti-rotational TPLO reduction pin in relation to tibial tuberosity fractures has been studied, no study to date has evaluated if a TPLO reduction pin left in situ will have a protective effect on the tibial tuberosity fractures during the initial 8 weeks of healing .The objective of this study was to compare the relative risk of tibial tuberosity fractures from TPLO procedures using a TPLO reduction pin in situ versus patients with a TPLO reduction pin removed at the time of surgery. Our hypothesis is that patients with a TPLO reduction pin left in situ will have a decreased incidence of tibial tuberosity fractures.Medical records for dogs treated with TPLO surgeries from January 2011 to May 2017 that had a TPLO surgery were reviewed. Dogs were included if a TPLO was performed with at least three sets of properly positioned orthogonal radiographs from immediately pre-operatively, post-operatively and at least 8 weeks post-operatively. Dogs were excluded if concurrent procedures were performed on the tibia . No bilateral single session TPLO surgeries were performed during this time period. All second side TPLO surgeries were performed at least 8 weeks post-operatively following radiographic confirmation of a healed osteotomy. All TPLO surgeries included were performed by ACVS board-certified surgeons who had more than 4 years of experience in performing the TPLO.Records were selected chronologically and sequentially for patients with an in situ reduction pin (group 1) and without an in situ reduction pin (group 2) until 200 stifles were indentified in each group. For each patient included in the study, we recorded the weight, age at the time of surgery, implants used and complications up to the 8 week recheck examination.TPLO surgeries were all performed with a locking TPLO plate  in the following four sizes: 3.5 mm broad, 3.5 mm regular, 3.5 mm mini, or 2.7 mm. In the proximal segment locking cortical bone screws were used for all available locking holes. In the distal segment non-locking cortical bone screws were used for every surgery. A 5/64″ positive profile end threaded stainless steel pin  was used as the TPLO reduction pin as seen in Fig. The craniocaudal and mediolateral radiographs were reviewed at all the three time periods . Pre-operative radiographs were evaluated for TPA and maximal distance from the tibial tuberosity to the caudal tibial plateau. Immediate postoperative radiographs were evaluated for TPA and the narrowest width of the tibial crest , defined as the width at the narrowest point of the tibial crest cranial to the osteotomy and distal to the insertion of the patellar ligament on the tibial tuberosity. The relative tibial tuberosity width was obtained by dividing the absolute tibial tuberosity width by the craniocaudal width of the tibia as previously defined by Bergh et al. . This waDescriptive statistics were calculated for all dogs enrolled in the study and are presented in Table A total of 400 dogs were included in the study that fit the criteria of 200 consecutive TPLO surgeries performed with each group were included in the study. Group 1 had 6 male intact, 76 male neutered, 3 female intact, 115 female spayed. Group 2 had 96 male neutered, 3 male intact, 101 female spayed, 0 female intact. The mean age during surgery was 4.8 years for group 1 and 5.2 years for group 2. The most common breeds were mixed breed (MBD) (37.5%), Lab (21%), and Pitbull (7.5%). Fractures occurred in English BD, Boxer, Lab (2), MBD, Beagle, and a Rottweiler. The median weight for group 1 was 29.65 kg (IQR: 24.58–37.02) and group 2 was 31.50 kg (IQR: 24.95–39.00). Additional details of groups 1 and 2 are presented in Table The 3.5 mm TPLO locking plate was the most common plate used by each group. Group 1 had 129 (64.5%) 3.5 mm plates, 31 (15.5%) 3.5 mm mini plates 20 (10.0%) 3.5 mm broad plates, and 20 (10.0%) 2.7 mm plates used. Group 2 had 141 (70.5%) 3.5 mm plates, 27 (13.5%) 3.5 mm mini plates, 17 (8.5%) 3.5 mm broad plates, and 15 (7.5%) 2.7 mm plates used. The TPLO reduction pin used for all 3.5 and 3.5 mm broad plates was a 5/64″ end threaded. The 0.062″ end threaded TPLO reduction pin was used for all 2.7 mm plates.Based on the Student’s t-test there was a statistically significant difference in fractures observed in group 1 and group 2 with a significance level of p < 0.05. In univariate logistic regression analysis, only the covariate for the presence of the reduction pin was associated with a statistically significant reduction in the likelihood of tibial tuberosity avulsion [OR: 0.138 (95% CI 0.007–0.789)]. In the multivariate model, adjusting for potentially confounding covariates, the presence of the reduction pin was associated with an approximate 92% reduction in the likelihood of tibial tuberosity avulsion [OR: 0.075 ]. Full results of the multivariate model are presented in Table In group 1, one fracture was observed in 200 consecutive TPLO surgeries (0.5%). The single fracture occurred in a 4-year-old Rottweiler with a pre-op TPA slope of 38° and post-op TPA slope of 9°. The absolute tibial tuberosity width was 0.76 cm and relative tibial tuberosity width was 0.16. The tibial tuberosity fracture was slightly displaced (less than 3 mm) with evidence of caudal rock back of the proximal segment.In group 2 seven fractures were observed in 200 TPLO surgeries (3.5%). The mean age was 5 years, with a mean pre-op TPA of 30.2° and mean post-op TPA of 7.3°. The mean absolute tibial tuberosity width was 0.76 cm and relative tibial tuberosity width was 0.18.To our knowledge this is the only study of TPLO procedures that evaluates the incidence of tibial tuberosity avulsion fractures in relation to the presence of an in situ TPLO reduction pin. Our data reveals a significant statistical difference between a TPLO performed with an in situ reduction pin and without the pin. No complications were reported due to the presence of the reduction pin in the time frame studied.Tibial tuberosity fractures of 3.5% reported in group 2 (no pin) were in line with the previously reported tibial tuberosity fractures of 1–9% for TPLO’s performed without an in situ reduction pin . In the The correct anatomic placement of the TPLO reduction pin with regard to the tibial crest and the effect on tibial tuberosity fractures has been previously reported. In this study, our only tibial tuberosity fracture with the in situ reduction pin occurred when the pin was improperly placed distal to Sharpey’s fibers. Placement of the pin distal to Sharpey’s fibers can weaken the tibial tuberosity and cause a stress riser . Of the Based on our results, we can conclude that an in situ TPLO reduction pin does decrease the incidence of tibial tuberosity fractures in our studied population. Although there are multiple factors associated with tibial tuberosity fractures, such as pin location and TPA, we believe the inclusion of a properly placed in situ TPLO reduction pin may be a simple technique that the surgeon can incorporate to decrease the incidence of tibial tuberosity fractures.Limitations of this paper include the retrospective nature of the study and the potential selection bias. We attempted to limit selection bias by including all consecutive TPLO surgeries performed by one of the four board-certified surgeons and not limiting to a particular surgeon or particular size patient. It is possible that within our follow-up time tibial tuberosity fractures could have occurred and the owners elected care with their primary veterinarian or did not seek veterinary advice. In our experience, we have found that owners who have complications with a TPLO are more likely to follow-up with our hospital. Previous reports have found that tibial tuberosity fractures most likely occur before 5 weeks . While iAdditional file 1: Figure S1. Boxplot showing age (years) by avulsion status. Figure S2. Boxplot of avulsion status by dog weight. Figure S3. Boxplot of avulsion status by the difference in pre- and post-op angle.


Trichinella spp. encapsulating species. The main biological functions of NC were identified as hypermitogenic growth arrest and pro-inflammatory phenotype, both inferred to depend on AP-1 (activator protein 1) transcription factor. Since those functions, as well as AP-1 activity, are known to be regulated among other pathways, also by Wnt (Wingless-Type of Mouse Mammary Tumor Virus Integration Site) signaling, transcription profiling of molecules participating in Wnt signaling cascades in NC, was performed.The nurse cell (NC) constitutes in mammalian skeletal muscles a confined intracellular niche to support the metabolic needs of muscle larvae of 2 Profiler PCR Arrays and complemented by that obtained by searching microarray data sets characterizing NC transcriptome.Wnt signaling-involved gene expression level was measured by quantitative RT-PCR approach with the use of Qiagen RTThe genes involved in inhibition of canonical Wnt/β-catenin signaling cascade as well as leading to β-catenin degradation were found expressed in NC at high level, indicating inhibition of this cascade activity. High expression in NC of genes transmitting the signal of Wnt non-canonical signaling cascades leading to activation of AP-1 transcription factor, points to predominant role of non-canonical Wnt signaling in a long term maintenance of NC biological functions.Canonical Wnt/β-catenin signaling cascade is postulated to play a role at the early stages of NC formation when muscle regeneration process is triggered. Following mis-differentiation of infected myofiber and setting of NC functional specificity, are inferred to be controlled among other pathways, by Wnt non-canonical signaling cascades.The online version of this article (doi:10.1186/s13071-016-1770-4) contains supplementary material, which is available to authorized users. Trichinella spp. Its basic morphological structure, called cyst, is formed within mammalian striated muscles 20–28 days post-oral infection [1-like type accompanied by cellular senescence [The nurse cell (NC) constitutes an intracellular niche for the muscle larvae of parasitic nematode nfection , 2. Larvnfection . NC is cnfection , 4. Basenescence . NC was nescence .2+, signaling cascades [Wnt signaling pathway plays an important role in morphogenesis and postnatal stem cell fate determination , 8. Inhicascades , 16. Parcascades , 18. Descascades . Of notecascades . In the cascades .As far as skeletal muscles are concerned, Wnt signaling is involved in myogenesis and muscle regeneration. Canonical Wnt/β-catenin signaling mediated by Wnt 1 and Wnt 7A ligands was shown to induce early myogenesis in mice . Wnt 3A,As a cellular system, NC originates from muscle cells suspended during regeneration. Immunological activities with signaling pathways culminating at AP-1 transcription factor activation, were identified as its prominent biological functions . Those cTrichinella spiralis H2 human isolate, was exploited as previously described [Trichinellosis in BALB/c mice, infected with escribed . NCs werescribed . NC in a2 First Strand kit was used for reverse transcription. RT2 SYBR Green/ROX qPCR Master Mix was used for quantitative PCR on Qiagen RT2 Profiler PCR Array of Mouse Wnt Signaling Pathway. The run was performed on 7500 Sequence Detection System (Applied Biosystems), including all control reactions recommended by the arrays’ manufacturer. Target gene expression level was calculated according to Qiagen RT2 Profiler PCR Array handbook, applying the comparative threshold cycle (CT) method, with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) used as a reference gene. It is given as 2exp-ΔCT (± average deviation for n = 2), where ΔCT is CT (target gene)-CT (GAPDH). The genes included into RT2 Profiler PCR Array whose threshold cycle fell above 35th cycle were excluded from data presentation.Qiagen kits were used at all steps. Total RNA was isolated with the use of RNaesy Mini kit, according to manufacturer’s instruction, with implementation of the RNase-Free DNase digestion step. RNA integrity was confirmed using Agilent 2100 BioAnalyzer. RT2 Profiler PCR Array, as well as the identifiers not included in the PCR array but otherwise related to Wnt signaling. In the aforementioned competitive microarray analysis, the transcriptomes of C2C12 myoblasts and C2C12 myotubes served as referral systems to the NC transcriptome. In order to eliminate biological differences among NC preparations, four different preparations of NCs isolated from mice carrying 5 to 12 month-infections were exploited for competitive microarray analysis. Only the identifiers with differential gene expression level ≥ 2 accompanied by a P-value ≤ 0.05, were considered signaling pathway-eligible. Fold change in gene expression level in NC, in relation to C2C12 myoblasts or myotubes, was calculated form log2ratio value and is provided as the average of quadruplicates, accompanied by the P-values calculated by Student’s one-sample t-test. All parameters of statistical analysis, including log2ratio ± standard deviation (SD) as well as the t-values, are shown in Additional file In order to complement performed herein transcription profiling of Wnt signaling factors in NC, the previously obtained competitive microarray data sets , were sevs uninfected muscle tissue [The NC is a non-muscular structure originating from a few types of cells. During encapsulation of the larva lasting up to 28 days post-infection, the nuclei, mitochondria and basophilic cytoplasm (i.e. staining with haematoxylin in haematoxylin and eosin (H&E) staining protocol), of infected myofiber, degenerate with the signs of apoptosis and autocrine signaling by tumor necrosis factor α , 25. Inhe tissue . Wnt cane tissue , was alse tissue . These fA network of molecules participating in Wnt signaling, whose expression was detected in NC, is depicted in Fig. It is thus inferred that dominant expression in NC of molecules involved in β-catenin degradation as well as inhibition of canonical Wnt signal transduction and β-catenin-dependent transcription, indicate that even though could be operating, Wnt/β-catenin signaling cascade is inhibited. Expression of the cascade effector genes may have resulted from Wnt/β-catenin-activated transcription at the earlier stages of NC formation, but in fully established NC this regulation seems to be attributed rather to other signaling pathways.2+ and protein kinase C axis, known to be activated also by classical GPCRs. JNK3 phosphorylates c-Jun and JunD which then dimerise with one of the Fos proteins to form transcription factor AP-1, known to display prosurvival and proinflammatory action, as well as to inhibit myogenesis [2+ signaling cascade, expressed in NC, is Nemo-like kinase (NLK, Table A network of molecules participating in non-canonical Wnt signaling cascades, whose expression was detected in NC, is schematically depicted in Fig. ogenesis , 42–47. It is inferred from the study performed that canonical, as well as non-canonical cascades operate in NC at the various stages of its formation. Expression in the fully established NC of Wnt ligands responsible for activation of canonical Wnt signaling cascade, the latter known to accompany induction of muscle regeneration , 22, mayTrichinella spp. muscle larvae. Assuming autocrine signaling by Wnt ligands to occur during a long-term existence of the NC-Trichinella muscle larva complex, the canonical Wnt signaling cascade is inferred to be inhibited, but the non-canonical Wnt/PCP and Wnt/Ca2+ cascades are postulated to lead to maintenance of AP-1 transcription factor activation and execution of NC biological functions.The NC is an intracellular habitat for


While the mortality rates for cervical cancer have been drastically reduced after the introduction of the Pap smear test, it still is one of the leading causes of death in women worldwide. Additionally, studies that appropriately evaluate the risk of developing cervical lesions are needed. Therefore, we investigated whether intracellular signaling entropy, which is measured with microarray data, could be useful for predicting the risks of developing cervical lesions. We used three datasets, GSE63514 (histology), GSE27678 (cytology) and GSE75132 . From the data in GSE63514, the entropy rate was significantly increased with disease progression  . From the data in GSE27678, similar results  were obtained . From the data in GSE75132, the entropy rate tended to be higher in the HPV-persistent groups than the HPV-negative group. The group that was destined to progress to CIN 3 or higher had a tendency to have a higher entropy rate than the HPV16-positive without progression group. In conclusion, signaling entropy was suggested to be different for different lesion statuses and could be a useful biomarker for predicting the development of cervical intraepithelial neoplasia. Cervical lesions are classified into cervical intraepithelial neoplasia (CIN) 1, 2 and 3 based on its relationship with the prognosis. CIN 1 is mild dysplasia, which is mostly observed because it disappears as part of its natural course. CIN 3 includes severe dysplasia and carcinoma in situ, and management involves treatment because it is highly likely to develop into invasive cancer. CIN 2 is moderate dysplasia, and the choice of whether to observe or treat the patient depends on the patient’s condition and wishes. Generally, conization, which is one of the most common treatments for cervical lesions, increases the risks of postoperative complications and pregnancy complications, such as abortion and preterm labor [While the mortality rates of cervical cancer have been drastically reduced since the introduction of the Pap smear test, it remains one of the leading causes of death in women worldwide . The defrm labor , 5. Thusrm labor . AlthougMeanwhile, there have been many investigations on the diversity or heterogeneity of various solid tumor types , 8. The Taken together, we aimed to investigate whether measuring signaling entropy was useful for both distinguishing cervical cancer cells from normal cells and predicting the risk of developing cervical precancerous lesions. In other words, we hypothesized that signaling entropy in cervical lesions would increase with the disease progression. In addition, we also hypothesized that signaling entropy should help discriminate the pathologically normal cells if their prognoses differ.Indeed, we confirmed that the signaling entropy increased with disease progression. Additionally, we found that the signaling entropy increased with HPV infection even in cells that were pathologically normal.In the present study, we investigated whether measuring the signaling entropy can be useful for predicting the risks of developing cervical lesions. Further investigation is needed to determine how signaling entropy changes in each patient; however, signaling entropy was suggested to be a useful biomarker for predicting the development of cervical intraepithelial neoplasia.www.sourceforge.net/projects/signalentropy [In this study, a Protein Interaction Network (PIN) was constructed, and the signaling entropy (SR) was measured as described in the literature , 10. Brilentropy , 10. If lentropy , 10.The datasets used in this study were GSE63514, GSE27678 and GSE75132 , 16. We GSE63514 contained 128 samples . The platform used was an Affymetrix Human Genome U133 Plus 2.0 Array. The data were already RMA-normalized, and 19151 genes were annotated to Entrez Gene ID. Among them, 8127 genes were included in the PIN in the program.GSE27678 contained 77 samples . The platforms used were an Affymetrix Human Genome U133 Plus 2.0 Array and Affymetrix Human Genome U133A 2.0 Array. The data were already RMA-normalized, and 12460 genes in common were annotated to Entrez Gene ID. Among them, 6932 genes were included in the PIN in the program.GSE75132 contained 41 samples . The platform used was an Affymetrix Human Genome U133 Plus 2.0 Array. The data were not normalized, and RMA-normalization was performed. Here, 19150 genes were annotated to Entrez Gene ID. Among them, 8127 genes were included in the PIN in the program.p values less than 0.05 were considered statistically significant. Wilcoxon rank-sum test was used for comparing medians between two groups. Kruskal-Wallis test was used for comparing more than two groups.For statistical analysis, JMP Pro 11 was used, and GSE63514 was from the Study to Understand Cervical Cancer Early Endpoints and Determinants (SUCCEED study), and it contained 128 samples [Capture Microdissection System was then used to capture the epithelial lining of the cervix from normal healthy control specimens or precancerous and invasive cancerous cell masses from cervical lesions. Afterwards, the specimens were processed for comprehensive human mRNA-level measurements. As shown in We then investigated whether the above results could be applied to the data from cytological specimens. Cytology tests are used for screening, and, as a result, low-grade squamous intraepithelial lesions (LSILs) are likely to match CIN 1 in biopsies and high-grade squamous intraepithelial lesions (HSILs) are likely to match CIN 2–3 in biopsies. GSE27678 contained microarray data from 75 samples  and 2 cell lines  . The plain situ (including adenocarcinoma in situ), CIN3 or cancer (including adenocarcinoma). Finally, the dataset contained 41 samples . The platform used was an Affymetrix Human Genome U133 Plus 2.0 Array. As shown in Thus far, we observed the distribution patterns of the entropy rate in lesions. We then investigated whether the entropy rate could be a biomarker for predicting the prognosis of cervical lesions. GSE75132 was a dataset from a prospective cohort study . EnrolleIn the present study, we showed that the signaling entropy could be useful for predicting the prognosis of cervical epithelia.in situ and invasive cancer. Therefore, many researchers have investigated how to appropriately evaluate the risk of developing cervical lesions. We sought to detect high-risk cases by focusing on signaling entropy.Cervical cancer is one of the leading causes of death in women worldwide . The cauThere are multiple reasons why we focused on signaling entropy, especially in cervical cancer. In the literature, authors have proposed a measure of signaling pathway promiscuity as an approach for quantifying the stemness and heterogeneity of any given cancer sample . In otheIndeed, we confirmed that the signaling entropy increased along with the disease progression. Additionally, we found that the signaling entropy could help identify the prognosis of cells that were pathologically normal. In addition, the signaling entropy was shown to be able to help predict the prognosis of cervical epithelia.There are two limitations in the present study. One is that the ranges of the signaling entropy  were different in each experiment. This is thought to be due to, at least in part, the differences in the experimental procedures, such as biopsies and cytologies, and the methods for collecting specimens in an individual facility, the numbers and kinds of cells collected, and the platforms used for the microarray. Therefore, we could not investigate cut-off values of the signaling entropy. The other limitation is that GSE63514 and GSE27678 were datasets from retrospective studies, and GSE75132 was a dataset in which the signaling entropy could only be measured at one point even though it was from a prospective study. In other words, we could not identify the time course of signaling entropy in each patient. For instance, we do not know whether a CIN 2 patient with a high signaling entropy rate is likely to progress to CIN 3 or if a CIN 2 patient with a low entropy rate is likely to regress to CIN 1. We also do not know whether the signaling entropy gradually increases or drastically increases . Signaling entropy is known to be more increased by inactivation of tumor suppressor genes than activation of oncogenes –24. CollThe strength of measuring the signaling entropy is that this approach comprehensively assesses the data. While omics data analysis enables us to obtain substantial information, it also detects variables of uncertain significance . Thus, aIn the present study, we investigated whether measuring the signaling entropy can be useful for predicting the prognosis of cervical epithelia. Further investigation is needed to determine the time course of the signaling entropy in each patient; however, the signaling entropy was suggested to be a useful biomarker for predicting the development of cervical intraepithelial neoplasia.


Plasmodium falciparum malaria can trigger intervillositis, a local inflammatory response more strongly associated with low birthweight than placental malaria infection alone. Fetal growth (and therefore birthweight) is dependent on placental amino acid transport, which is impaired in placental malaria-associated intervillositis. Here, we tested the hypothesis that mechanistic target of rapamycin (mTOR) signaling, a pathway known to regulate amino acid transport, is inhibited in placental malaria-associated intervillositis, contributing to lower birthweight.Placental We determined the link between intervillositis, mTOR signaling activity, and amino acid uptake in tissue biopsies from both uninfected placentas and malaria-infected placentas with and without intervillositis, and in an in vitro model using primary human trophoblast (PHT) cells.We demonstrated that (1) placental mTOR activity is lower in cases of placental malaria with intervillositis, (2) placental mTOR activity is negatively correlated with the degree of inflammation, and (3) inhibition of placental mTOR activity is associated with reduced placental amino acid uptake and lower birthweight. In PHT cells, we showed that (1) inhibition of mTOR signaling is a mechanistic link between placental malaria-associated intervillositis and decreased amino acid uptake and (2) constitutive mTOR activation partially restores amino acid uptake.Our data support the concept that inhibition of placental mTOR signaling constitutes a mechanistic link between placental malaria-associated intervillositis and decreased amino acid uptake, which may contribute to lower birthweight. Restoring placental mTOR signaling in placental malaria may increase birthweight and improve neonatal survival, representing a new potential therapeutic approach.The online version of this article (doi:10.1186/s12916-016-0759-3) contains supplementary material, which is available to authorized users. Plasmodium falciparum malaria still affects about 85 million pregnancies each year [One in six infants worldwide are born with low birthweight <2500 g), which is the main risk factor underlying 80% of neonatal deaths . The Wor500 g, whach year . Little P. falciparum-infected erythrocytes in the maternal intervillous blood space of the placenta. This can trigger the recruitment and activation of maternal immune cells, resulting in a local inflammatory response termed intervillositis. Placental malaria-associated intervillositis is more strongly associated with low birthweight than placental malaria without intervillositis [Malaria in pregnancy can lead to placental malaria characterized by the sequestration of llositis . The undllositis , 6. TherPlacental nutrient transfer controls fetal nutrient availability, which is a key determinant of fetal growth (and therefore birthweight). The placental capacity to transfer nutrients is highly dependent on the expression and function of nutrient transporters in the syncytiotrophoblast (the nutrient transporting epithelium of the human placenta) . System The mechanistic target of rapamycin (mTOR) signaling pathway is a nutrient-sensing pathway that regulates cell growth, proliferation, and metabolism in response to hormones, growth factors, and nutrient availability . Primary human trophoblast (PHT) cells were isolated by trypsin digestion and Percoll centrifugation as originally described  with mod+ cells were cultured in 1:1 mixture of Dulbecco’s modified Eagle’s medium  and Ham’s F-12 nutrient mixture (Invitrogen) supplemented with 10% fetal bovine serum and with penicillin/streptomycin.Conditioned media were prepared as previously described  with modPHT cells were transfected with 10 nM DEPTOR small interfering RNA  or with Scramble siRNA control using Lipofectamine RNAiMax transfection reagent (Thermo Scientific) according to the manufacturer’s protocol.+-dependent uptake of 14C-methyl-aminoisobutyric acid (MeAIB) as previously described [System A activity was assessed by measuring the Naescribed .37/46), anti-ribosomal protein S6, anti-phospho-ribosomal protein S6 (Ser235/236), anti-Akt, anti-phospho-Akt (Ser473), and mouse anti-β-actin (Sigma-Aldrich). Horseradish peroxidase-conjugated secondary anti-rabbit and anti-mouse antibodies  were used as secondary antibodies. Proteins were visualized using chemiluminescence detection. Densitometry was performed using NIH’s ImageJ software.Placental homogenates and PHT cell lysates were loaded and proteins separated on 4–12% Bis-Tris gels (Invitrogen) or 12% Mini-Protean protein gels (Bio-Rad) and transferred onto 0.20-μm nitrocellulose membrane  or polyvinylidene fluoride membrane (Bio-Rad). Membranes were incubated with primary antibodies: rabbit anti-4EBP-1, anti-phospho-4EBP-1  according to manufacturer’s instructions.Data are presented as medians and scatter plots or medians and inter-quartile range. Data were analyzed and graphs designed using Prism 5 software (Graph Pad). Two-group comparisons were made using Mann–Whitney test and three-group comparisons using Kruskal–Wallis test. Spearman’s correlation test was used to assess correlations with 95% confidence interval.P. falciparum infection and intervillositis on mTOR signaling in human placenta using well-established mTOR functional readouts as summarized in Additional file Ser235/236 and 4E-BP1Thr37/46) and mTORC2 (AktSer473)  and to infected placentas without intervillositis for Akt signaling (Akt: P = 0.02). These results are consistent with a causal link between placental malaria-associated intervillositis and inhibition of mTOR signaling. This was further supported by the negative correlation between mTOR signaling activity and the severity of intervillositis measured as the percentage of monocytes in the intervillous space (Table We aimed to assess the impact of 73) Fig. . The act73) Fig.  was lowen = 38; r = −0.27; P = 0.093). Collectively, these data are consistent with the model that placental malaria-associated intervillositis decreases placental mTOR signaling, which reduces amino acid uptake and birthweight.Inhibition of mTOR signaling may lead to reduced birthweight by impacting amino acid transport. We determined the relationship between placental mTOR signaling activity, amino acid transport, and birthweight. mTOR signaling showed a positive correlation with our previous microvillous plasma membrane amino acid uptake data  measuredP. falciparum-infected erythrocyte co-culture (herein “infected conditioned medium”). This conditioned medium displayed high levels of inflammatory cytokines such as IL-1β, IL-6, IL-8, TNF, and IL-10  or to control medium   or to control medium (P = 0.03)  Fig. . The viaEBP-1: 03P = 0.004)  Fig.  without P = 0.004). This suggests that placental mTOR activation could partially restore placental amino acid uptake in response to placenta malaria-associated intervillositis.We quantified the decrease in System A activity induced by the infected conditioned medium in PHT cells transfected with DEPTOR siRNA versus PHT cells transfected with Scramble siRNA. In cells transfected with Scramble siRNA, System A activity decreased by 72% in response to the infected conditioned medium whereas System A activity in DEPTOR-silenced cells was decreased by only 50% Fig. . This deReducing the incidence of low birthweight remains a global health priority to minimize neonatal morbidity and mortality, impaired infant growth and cognitive development, and chronic diseases later in life . UnderstThe mTOR signaling pathway has been proposed to play a central role in placental nutrient sensing . This moP. falciparum-infected erythrocytes. This conditioned medium displayed high levels of IL-1β, IL-6, IL-8, TNF, and IL-10. Cytokines such as IL-1β have been shown to inhibit mTOR activity [Our ex vivo results reported here and in our previous publication  have demactivity  and simiactivity , 29. ThemTORC1 is the master regulator of the translational machinery and activates protein synthesis by phosphorylating downstream targets including 4EBP-1  stress inhibits Akt phosphorylation , 32 and We established mTOR signaling as a mechanistic link between placental malaria-associated intervillositis and reduced amino acid uptake using RNAi-mediated silencing of DEPTOR, an endogenous mTOR inhibitor . KnockdoOur results provide evidence for a causal link between placental malaria-associated intervillositis, mTOR signaling inhibition, and decreased amino acid uptake, an important determinant of fetal amino acid availability and fetal growth. The positive correlation between mTOR signaling and birthweight further supports the hypothesis that placental mTOR signaling influences fetal growth. We chose to focus on System A because this amino acid transporter is regulated by mTOR and is the placental transporter system most strongly associated with fetal growth . FurtherOur findings identify placental mTOR as a potentially valuable target for interventions aimed at improving birthweight in malaria in pregnancy. There is strong interest in targeting mTOR activity in therapeutic development and various mTOR signaling regulators have recently been tested in clinical trials for other conditions . Some arVarious malaria control measures have decreased malaria prevalence but with limited impact on the birthweight of babies born to malaria-infected women. Similarly, modest gains in birthweight have been achieved using untargeted maternal dietary supplementation , 6. TherAdditional file 1: Supplementary materials and methods, Table S1, and supplementary figure legends. (DOC 113 kb)Additional file 2: Figure S1.Correlation between mTOR signaling activity and the degree of intervillositis, System A activity, and birthweight. (TIFF 491 kb)Additional file 3: Figure S2.Cytokine profiles in conditioned media. (TIF 191 kb)Additional file 4: Figure S3.Syncytialization and viability of cultured primary human trophoblasts. (TIF 498 kb)


Salvador is the city with the highest number of HTLV-1 infected individuals in Brazil, yet the main route of HTLV-1 transmission is unknown.To investigate the association of syphilis infection as a proxy for sexual transmission of HTLV-1 infection in the general population of this city.Treponema pallidum infection was determined by serology. Logistic regression analysis was used to evaluate the independent effect of the potential explanatory variables to HTLV-1 infection and Odds Ratios (OR) and 95% CI were calculated.A cross sectional population-based study was conducted with 3,451 serum samples obtained by a representative simple random sampling. Data on gender, age, income, and years of education were collected by questionnaire and the presence of HTLV, HIV and ADJUSTED 36.77; 95% CI 14.96–90.41).The majority of studied individuals were female (56.4%), had less than 7 years of education (55.3%) and earned two or less minimum wages (52.0%). The overall prevalence of HTLV-1 was 1.48% , which increased with age. Only three persons younger than 17  years were infected by HTLV-1. Among the 45 syphilis positives, 12 (26.7%) were HTLV positive, while among 21 HIV positives, only one (4.8%) was HTLV positive. HTLV-1 infection was found to be associated with syphilis infection (ORThe data presented herein indicate that horizontal transmission between adults is the main route of HTLV-1 infection in the general population of Salvador and that this is likely to occur through sexual contact. Human T lymphotropic virus type-1 (HTLV-1) is a delta retrovirus of worldwide distribution and it is estimated that at least 5–10 million people harbor the virus . The higHTLV-1 is etiologically linked with adult T cell leukemia-lymphoma (ATLL) , tropicaThe inter-human transmission of HTLV-1 arises from infected lymphocytes, as plasma is unable to promote infection  and HTLVSalvador, located in the northeastern region of Brazil, is considered an epicenter of HTLV-1 infection in the country, with a prevalence above 1% in the general population , 1.35% , and confirmed by indirect immunofluorescence .Sera from all volunteers were screened for HTLV 1/2 antibodies by ELISA (HTLV-1 (rp21e enhanced), EIA, Cambridge Biotech Corporation, Worcester, MA, USA); confirmation and discrimination between HTLV-1 and HTLV-2 was performed by western blotting . HIV antibodies were detected using ELISA , and confirmed by indirect immunofluorescence , following the manufacturer’s recommendations. The outcome variable was prevalence of HTLV-1 infection. Syphilis infection was used as a proxy for sexual transmission of HTLV-1 infection, since other sexual behavior variables were not available in the main survey. Descriptive statistics were determined for each variable. Age was examined in four different strata to analyze trends in HTLV-1 prevalence rates . Other variables (and cut points) were years of education (>7 years); income (<twice the Brazilian minimum monthly wage (MW), equal to approximately US $125 at the time of survey. Prevalence rates of HTLV-1 infection and respective binomial 95% confidence intervals (CI) pertaining to study variables were estimated. Odds Ratios (OR) and 95% CI from logistic regression analyses were used to evaluate the association of syphilis infection with HTLV-1 infection. The Statistical Package for the Social Science  version 14.0 was used for statistical analyses.The study population comprised 3,451 subjects of which 1,947 (56.4%) were females; age ranged from 0 to 99 years. Eleven (0.32%) and 23 (0.67%) subjects provided no information about sex and age, respectively. The majority  had less than seven years of education and 52.0% (1660/3192) earned two MW or less.The overall prevalence of HTLV-1 was 1.48% , which was shown to increase with age . HTLV-1 crude: 44.63; CI95%: 6.17–116.79).HTLV-1 infection was strongly associated with syphilis seroreactivity  after adjusting for other study variables . Males pThe city of Salvador continues to be an important location for HTLV-1 infection surveillance, as the prevalence among the general population remains above 1% (1.45%). In addition, very few studies involving large samples have investigated HTLV among the general population .T. pallidum seropositivity were almost 40 times more likely to be HTLV-1 positive compared to T. pallidum soronegative individuals. Second, the seroprevalence of HTLV-1 infection in children and adolescents younger than 17 years was only 0.3%. These HTLV-1 infected persons were two girls, aged two and fourteen years old, and one 16-year-old boy. Third, a sharp increase in the prevalence of HTLV-1 infection in older age groups confirms the adult acquisition of HTLV-1, supporting the previously postulated predominance of sexual transmission [Three observations from this study strongly suggest that sexual contact constitutes the main route of HTLV-1 transmission in the general population of Salvador. First, a robust association between syphilis infection and HTLV-1 infection. Those with smission .Unprotected sex, multiple partners, history of other sexually transmitted diseases, and sexual intercourse with injection drug users were identified as risk factors for HTLV-1 infection in this city , 28. In Evidence of sexual transmission as a main route of HTLV-1 has also been reported in the Caribbean , 31. ConAlthough sexual transmission was the most probable route of HTLV-1 infection in this study, vertical transmission also plays a role in the dissemination of HTLV-1. Previous studies have shown that the prevalence of HTLV-1 infection among pregnant women in Salvador and other cities in the state of Bahia remains around 1% , 38, 39.It has been postulated that a period of breastfeeding lasting six months or more is strongly associated with mother-to-child transmission . HoweverT. pallidum among individuals 17–30 years was 1.91%, nine times higher than that of younger individuals ≤ 16 years. Only two individuals aged 15 and 16 years had positive serology, which reinforces the notion that infection by T. pallidum should be considered a marker of sexual transmission. The higher prevalence in the general population could be explained by the failure of public health programs to control infection. For instance, a recently published editorial estimated that just 12% of sexual partners received treatment for syphilis in Brazil, which is clearly a failure by the public health system to control transmission of syphilis infection [In the present study, the prevalence of syphilis in the general population of Salvador was 1.3%, two times higher than the prevalence of HIV-1 (0.6%). Moreover, the prevalence of seropositivity for nfection . By contnfection  HIV trannfection –53.Preventive measures to avoid HIV and blood-borne infections may have also helped curb HTLV-1 infection in Brazil. However, there is a possibility that new prevention tools based on the treatment of HIV will not be applicable to the prevention of HTLV, potentially leading to higher rates of HTLV-1 transmission.The present study is limited due to the cross-sectional nature of the data herein; as such, we are unable to determine causal relationships of the observed associations between syphilis infection and HTLV-1. Nevertheless, the observed associations are consistent with previously presented results . In addiNonetheless, it is of paramount importance to reinforce preventive measures, such as urging the use of condoms, as well as to implement mandatory HTLV screening for pregnant women throughout Brazil.