Inheritance of the apoE4 allele is the strongest genetic risk factor for late-onset AD. However, there is no consensus on how different apoE isotypes contribute to AD pathogenesis. It has been hypothesized that apoE and apoE4 in particular is an amyloid catalyst or “pathological chaperone”. Alternatively it has been posited that apoE regulates Aβ clearance, with apoE4 been worse at this function compared to apoE3. These views seem fundamentally opposed. The former would indicate that removing apoE will reduce AD pathology, while the latter suggests increasing brain ApoE levels may be beneficial. Here we consider the scientific basis of these different models of apoE function and suggest that these seemingly opposing views can be reconciled. The optimal therapeutic target may be to inhibit the interaction of apoE with Aβ rather than altering apoE levels. Such an approach will not have detrimental effects on the many beneficial roles apoE plays in neurobiology. Furthermore, other Aβ binding proteins, including ACT and apo J can inhibit or promote Aβ oligomerization/polymerization depending on conditions and might be manipulated to effect AD treatment.The amyloid cascade hypothesis remains a robust model of AD neurodegeneration. However, amyloid deposits contain proteins besides A The identification of mutations within the amyloid precursor protein (APP) and presenilin (PS) genes that cause autosomal dominantly inherited AD and that result in increased production of amyloid-prone forms of Apothesis , 2.β peptide, some forms of which have been termed ADDLs (Aβ-derived diffusible ligands), protofibrils, or Aβ*56 [β oligomers are not only potential intermediates in the formation of amyloid filaments, but they also have been shown to be neurotoxic themselves and to inhibit long-term potentiation (LTP), a cellular model of memory, in hippocampal slices [β oligomers in the neurodegeneration process.The role of amyloid in neuronal dysfunction has recently been extended by the discovery of small, soluble, oligomers of the Aor Aβ*56 –6. Thesel slices , 7, 8. Tβ, amyloid deposits also contained the inflammation/acute phase protein α1-antichymotrypsin (ACT) [ Despite its strength, the Amyloid Cascade Hypothesis is incomplete without including the essential role of amyloid-associated inflammatory proteins. For example, biochemical and histological studies first showed that, in addition to Ain (ACT)  and, latin (ACT) , 11, whiin (ACT) , 11, 12.in (ACT) –15). Indin (ACT) . Howeverin (ACT) , 18.ε4 allele was found to be the strongest known risk factor for AD besides age, with one copy increasing AD risk 3–5-fold and two copies over 10-fold [ε4 promotes cognitive decline in middle-aged Down syndrome individuals [The significance of these biochemical results instigated and was reinforced by parallel genetic discoveries implicating a role for inflammation in AD. In particular, inheritance of the apoE  10-fold –21. Furtividuals .β and apoE and other inflammatory proteins, on the effects of such interactions, and on their implications for designing apoE-based AD therapies. The central question we try to answer is whether increasing or decreasing apoE level and/or function will serve best to reduce AD/DS pathology and cognitive decline. Lack of a clear answer may lead to the development of drugs that, rather than serving as an AD therapy, instead potentially exacerbate the disease.Because of apoE's essential genetic, and therefore presumably biochemical, contribution to AD pathology and cognitive decline, it is critical that its role in the AD pathogenic pathway/amyloid cascade be elucidated in order for therapeutics based on apoE to be designed. While recent excellent and encyclopedic literature reviews describe the many potential roles that apoE plays in AD –26, thisin vitro and in vivo studies showed that mature amyloid deposition and the associated cognitive decline is strongly stimulated by apoE and ACT in a dose-dependent and isoform-specific manner, with apoE4 being the strongest promoter of Aβ polymerization and apoE2 being an inhibitor, paralleling the effect of these two isoforms in humans [β expression equal to the apoE-expressing APP animals. Elegant work by Manelli and colleagues also showed that native lipidated apoE4 from transgene replacement astrocytes increases Aβ neurotoxicity compared to apoE3 or E2, indicating that apoE4 provides a negative gain of function [β as oligomers, the most neurotoxic form of the Alzheimer amyloid peptide [β during biochemical isolation of amyloid from human brains, and that apoE preferentially interacts with Aβ peptides in a β-sheet structure [To determine whether inflammation contributes to Alzheimer's disease rather than being merely a correlative pathological feature in the AD brain, we and others tested the hypothesis that ACT and/or apoE serve as amyloid catalysts or pathological chaperones. Numerous n humans –38. Indefunction . Finally peptide , 41. Thetructure –45.β monomer, and no AD would ensue.Together these results show that inflammatory proteins, particularly apoE, are integral parts of the amyloid cascade, and that without them the cascade would be arrested at the level of the harmless Aβ from the brain. Under this view, ApoE is protective, with human apoE4 being less protective than apoE3 or E2 .The view of apoE as an integral and pathological part of the amyloid cascade has been shaken by experiments that suggest that apoE, far from being an amyloid catalyst, serves to clear Aion, see  and commβ clearance from the brain compared to mouse apoE, with apoE4 inhibiting clearance the most. Other experiments showed that indeed, clearance of Aβ species was inhibited by complexing with apoE, especially apoE4 [The first experiments that suggested apoE's role as a neuroprotector examined the pathology and cognition of APP transgenic mice carrying a second transgene expressing one or another human apoE isoform. Contrary to expectations, amyloid deposition in these mice was inhibited by the human apoE transgene, as though human apoE was protective . Ultimatly apoE4 , 50.β clearance mechanism appeared to be supported by the finding that introduction of anti-Aβ antibodies or other Aβ-binding proteins such as gelsolin, led to a reduced amyloid load in the brain and rapidly improved cognition, with little evidence of Aβ-binding agents invading the brain parenchyma [β-binding peripheral protein.The possibility that interaction with apoE modulated an Arenchyma –54. We arenchyma . Thus thβ and improves cognition in APP Tg mice, while knocking out the ABCA1 gene in APP mice showed a tendency to reduced amyloid load. Because apoE expression and lipidation is stimulated by LXR activation, the results were interpreted as proof that increased apoE levels help microglia clear Aβ and amyloid, as indeed some earlier cell culture experiments had suggested. However, it has also been shown that genetic overexpression of ABCA1 reduces amyloid deposition in mice where the apoE levels are unchanged [in vivo action to the altered level of one particular protein. Furthermore, the increased levels of ABCA1 induced by Bexarotene enhance apoE lipidation, a change that is known to alter apoE/Aβ interactions. Hence, it is important to consider the lipidation state of apoE, which affects its function, in addition to the absolute levels of apoE.Most recently, an approach to therapy has been investigated in AD mice that is based on activating the liver X receptor (LXR), which also exists on other cells including microglia –57. Actinchanged . Hence, β clearance with human apoE4 being a greater inhibitor of clearance (or poorer clearer), then reducing apoE levels or apoE binding to Aβ should increase amyloid deposition and cognitive decline. When trying to distinguish and weigh the value of two hypotheses, it is instructive to consider their testable predictions. If apoE is an amyloid catalyst, then reducing apoE levels or function in the brain should result in reduced amyloid deposition and reduced cognitive decline. If on the other hand, apoE is involved in Ain vitro or in transgenic mice indicate that the ability of Aβ to form neurotoxic filaments or oligomers and cause cognitive decline are increased in the presence of apoE, particularly mouse apoE and human apoE4, with apoE2 being protective. In contrast, in the complete absence of apoE, the mutant APP gene and its product Aβ are harmless, generating neither amyloid deposits, synaptic disfunction, or cognitive decline, with one copy of apoE having an intermediate effect, as discussed above. The in vitro experiments in particular indicate that apoE likely acts catalytically to promote Aβ polymerization, as the molar ratio of Aβ to apoE of about 200/1 was appropriate for the formation of neurotoxic products [ All experiments carried out so far products –30. Mostproducts , 60.in vitro, cell culture, and transgenic mouse data is that apoE is necessary for Aβ to polymerize into neurotoxic oliogomers/filaments, probably by binding to Aβ and thus altering its structure more toward the β-sheet and more easily allowing successive Aβ peptides to add on to the growing chain. The recent finding that apoE promotes Aβ oligomer formation in vivo reinforces this interpretation [ The simplest interpretation of the retation , 41. Wheβ filament catalyst, have been borne out, the compelling experiments demonstrating that human apoE inhibits filament formation in a mouse background require explanation. Furthermore, data from LaDu and colleagues and by others have shown that lipidated apoE, presumably the prevalent form in vivo, binds Aβ with an affinity of E2 > E3 > E4 [β in the brain interstitial fluid compared to E2 or E3 [ Even though the key predictions of the polymerization hypothesis, that is apoE serving as an A E3 > E4 –64. FinaE2 or E3 .β binding studies might be interpreted as support for apoE functioning in Aβ clearance because apoE2, for example, would bind Aβ tightly and could thereby promote its removal from the interstitial fluid via LRP receptors [β most tightly, and thereby not only prevent apoE4 from binding and promoting Aβ oligo/polymerization, but also prevent the spontaneous polymerization of the peptide. The apoE-Aeceptors , 61–64. eceptors , 66. If β with different strengths can also explain why human apoE isoforms slow amyloid deposition in the presence of the endogenous mouse apoE, for they may bind Aβ more tightly or differently than mouse apoE and slow the catalytic conversion of Aβ into oligomers/polymers in the mouse background. The ability of different apoE isoforms to bind Aβ clearance can also be interpreted as reflecting apoE's role in catalyzing Aβ oligo/polymerization. Pathologic macromolecular structures are often resistant to various clearance mechanisms designed for monomeric species, whether by intracellular proteasome degradation or cross-membrane/BBB transfer, thus allowing their accumulation. Only when oligo/polymeric structures are anticipated and physiological clearance mechanisms are in place to handle them, as for antibody-antigen complexes, will clearance be facilitated by conversion to larger structures. Because apoE clearly has the ability to catalyze the conversion of Aβ into oligomeric and polymeric structures, it is reasonable to assume that those structures will be more difficult to clear, and that such difficulty will be detected as clearance inhibition in the brain, for instance, by apoE4, in pulse chase type experiments, while the higher apoE levels in blood may aid the clearance of Aβ from the circulations  and apoE (Aβ12-28) bind can serve as decoy peptides that prevent the binding of apoE to Aβ and its catalysis of Aβ into neurotoxic species [in vitro work has recently been repeated and confirmed in other laboratories [in vivo by preparing a version of Aβ12-28 that has a better plasma 1/2 life and is nonfibrillogenic/nontoxic. It was shown that this peptide could be peripherally introduced into a transgenic APP mouse, where it effectively entered the brain and prevented/reversed oligomer formation, amyloid deposition, and cognitive decline [β1-11 into one side of the APP mouse brain to block ACT's binding site with endogenous Aβ rapidly reduced amyloid load compared to the other, vehicle-injected side of the brain. Furthermore the inflammatory cytokine IL-1 that is overexpressed in AD brain [in vitro or in vivo, successfully prevents Aβ pathology and neurotoxicity. A good test of any hypothesis about the pathogenesis of a disease is whether it successfully predicts how the pathogenesis can be inhibited or reversed. For example, small fragments of A species . This earatories , 73. The decline –76. SimiAD brain  induces AD brain , and bloAD brain . Evidentβ and can be shown to aid its passage across the blood brain barrier [ Apolipoprotein J also binds A barrier –83. Inte barrier . This reβ binding proteins are specific anti-Aβ antibodies, which form the basis of both passive and active immunization therapies for Alzheimer's disease . The fiAPP mice . Evidentβ-binding proteins can have multiple effects on polymerization and that their full range of activities must be considered when using them as potential targets or tools for therapeutic intervention. These results illustrate the fact that Aβ oligomers have been shown to be highly neurotoxic in vitro and in vivo, and their formation is promoted by apoE4, the mechanism of their toxicity is still being elucidated. The data reviewed above coupled to other recent findings suggest a novel mechanism for Aβ toxicity that encompasses the essential role of apoE. Specifically, Aβ oligomers bind to and inhibit certain microtubule motors that are essential for the function and stability of the mitotic spindle—Eg5/kinesin5, Kif4A, and MCAK [β or by the specific kinesin 5 inhibitor Monastrol prevents the efficient transport of receptors such as the LDLR, the NMDA neurotransmitter receptor, and the p75 neurotrophin receptor to the cell surface, resulting in reduced function ([β into oligomers, which then inhibit MT-based transport of key cellular components such as receptors to their functional location. Although Aand MCAK . Similarand MCAK , 89. We unction (; in prepunction (. This laβ and catalyze its conversion into neurotoxic macromolecular species (in vivo demonstration that blocking apoE-Aβ interaction prevents AD in a mouse model, suggests that this decoy approach should be translatable into human patients and serve as an effective new approach to AD therapy. In sum, it appears that the preponderance of the data can be most consistently interpreted as showing that the brain inflammatory protein apoE plays a catalytic role in the AD/DS amyloid cascade and consequent cognitive decline, with binding and clearance differences between the apoE isoforms reflecting their differing abilities to bind to A species . This coβ-binding proteins may be similarly manipulated by a decoy approach to reduce oligomerization and polymerization of Aβ into neurotoxic species. However, the finding that different antibodies to Aβ can both inhibit ACT-catalyzed Aβ polymerization and promote polymerization of Aβ itself, argues that immunotherapy must be approached with care to avoid the use or induction of antibodies that can catalyze further oligo/polymerization of Aβ, instead of inducing its phagocytosis and removal. Furthermore, human and mouse intracerebral environments may differ in important ways with respect to the pattern and activities of Aβ-binding proteins and may also respond differently to intervention or inflammation. Such differences may explain why so many treatments that were successful in reducing amyloid-dependent cognitive decline in transgenic mice have failed to translate into human AD patients.Other Aβ oligomers to inhibit key microtubule motors and prevent the transport of neurotrophin, neurotransmitter, and other receptors to the cell surface may underlie their neuronal toxicity. It is apparently the ApoE-, especially E4-dependent formation of such Aβ oligomers, that constitutes the key catalytic step in the AD pathogenic pathway. Finally, the ability of A


However, the ε4 carriers fit with the general correlation pattern exhibited by the ε3/ε3 carriers, as ε4 carriers on average had longer telomeres and smaller hippocampi compared with ε3/ε3 carriers. The relationship observed can be interpreted as long telomeres representing a history of relatively low cellular proliferation, reflected in smaller hippocampal volumes. The results support the potential of leukocyte telomere length being used as a biomarker for tapping functional and structural processes of the aging brain.Telomere length shortens with cellular division, and leukocyte telomere length is used as a marker for systemic telomere length. The hippocampus hosts adult neurogenesis and is an important structure for episodic memory, and carriers of the apolipoprotein E ε4 allele exhibit higher hippocampal atrophy rates and differing telomere dynamics compared with non-carriers. The authors investigated whether leukocyte telomere length was associated with hippocampal volume in 57 cognitively intact subjects  aged 49–79 yr. Leukocyte telomere length correlated inversely with left ( Telomeres are stretches of repetitive non-coding sequences, constituting the ends of linear chromosomes. As cells divide, the process of chromosomal replication inevitably leads to telomere shortening due to the end-replication problem. If a critical length is reached, the cell will either undergo apoptosis or enter a state of replicative senescence APOE ε4 allele is a well-established genetic risk factor for Alzheimer's disease (AD) and influences episodic memory negatively in aging individuals, even in the absence of AD or mild cognitive impairment APOE ε4 carriers compared with non-ε4 carriers, and further that worse episodic memory was associated with longer leukocyte TL among ε4 carriers APOE ε4 allele. Smaller hippocampi among ε4 carriers have been most consistently found among AD patients, but have also been observed among cognitively healthy individuals Apolipoprotein E (APOE) is a polymorphic gene with three common alleles: ε2, ε3, and ε4, with typical population frequencies of approximately 5–10%, 70–85% and 10–25%, respectively. The APOE ε4 allele. The sample was drawn from the Betula study on memory and aging, located in northern Sweden, and consisted of 57 cognitively intact subjects, out of whom 29 carried the ε3/ε3 genotype, and 28 carried the ε4 allele.With the aforementioned as background, we set out to explore the potential relationship between hippocampal volume, as determined by magnetic resonance imaging (MRI), and leukocyte TL in both carriers and non-carriers of the p = 0.935). There were no significant differences between the 29 APOE ε3/ε3 and 28 APOE ε4 carriers regarding age, education, gender distribution, or any of the health parameters. Age-adjusted leukocyte TL was significantly longer among ε4 carriers , consistent with observations in the sample of which the current sample is part of p = 0.118). Hippocampus volume  among ε3/ε3 carriers showed a negative correlation with age , but there was no association between age and hippocampus volume among ε4 carriers (sr = −0.054 p = 0.783). Hippocampal volumes tended to be smaller among ε4 carriers, but the differences were not statistically significant , right  as well as total volume . No significant correlations between hippocampus volume and leukocyte TL were found among the ε4 carriers  and women (n = 18); indicating that both genders contributed to the observed correlation . Among ε4 carriers, no significant correlations were observed after stratifying for gender . Raising the MMSE inclusion criteria from ≥24 to ≥26, leading to exclusion of 5 ε3/ε3 and 5 ε4 carriers, did not alter the significances of the correlations . Considering the documented effect of age on both TL and hippocampal volume, age is a significant confounder in the analyses. Not taking the confounding effect of age and body/head size into account, i.e., performing the analyses completely unadjusted, weakened the correlations among ε3/ε3 carriers and made them non-significant ; showing that for TL to be significantly associated with hippocampal volume among ε3 homozygotes, age has to be taken into account . Analysis for further confounding did not reveal any confounding of the correlation between TL and hippocampus volume by any of the health parameters listed in Analyzing leukocyte TL (adjusted for age) and hippocampus volume (adjusted for age and body size) showed that among ε3/ε3 carriers, TL was significantly inversely correlated with hippocampus volume ; with leIn the present study, we investigated whether leukocyte TL had any relationship with hippocampus volume among 57 non-demented individuals, aged 49 to 79 yr. In our sample of 29 ε3/ε3 carriers, TL was inversely correlated with hippocampus volume. Among the 28 ε4 carriers, no correlation was present; although, as a group the ε4 carriers fit with the correlation pattern found among ε3/ε3 carriers as they in general had longer TL and smaller hippocampi, compared with ε3/ε3 carriers.APOE ε4 carriers, but not significantly. However, as shown by Lind et al. APOE ε4 in AD Hippocampal volumes tended to be smaller among APOE genotype into account, and included 7 (out of 26) participants with mild cognitive impairment (MCI). Considering that they also reported data showing that their participants with MCI had both smaller hippocampi and shorter TL (compared with the cognitively intact subjects), the inclusion of these subjects in their data analysis on the relationship between TL and hippocampal volume might to a large extent have explained their finding of a positive correlation. Our sample included no MCI cases, only cognitively healthy subjects.In contrast to our findings, Grodstein et al. APOE status) and they found their hippocampal telomeres to be on average 49% longer compared with hippocampal tissue from age-matched non-demented subjects. Although their paper did not contain any volumetric information regarding the hippocampi which were sampled, a consistent finding among AD patients is the exhibition of smaller hippocampi compared with those of age-matched controls Several studies that have compared TLs between different tissues in the same individuals have observed correlations between them; i.e., individuals with relatively long leukocyte TL have relatively long TLs in other tissues as well, although not necessarily of similar lengths in absolute terms Seeing as TL to some degree reflects historical proliferative activity, the finding by Thomas et al. TL was significantly longer among carriers of the APOE ε4 allele, as reported in our previous study APOE ε3/ε3 carriers, as well as part of the explanation as to why ε4 carriers as a group fit with the same inverse relationship. Low activity of proliferation can explain long telomeres coinciding with relatively small hippocampal volumes and fits with our previous findings linking long leukocyte TL with poor performance of episodic memory, a memory system believed to be dependent on adult neurogenesis taking place in the hippocampus. This hypothesis is also compatible with what was found by Thomas et al. In summary, we hypothesize that proliferative activity is the key to our observed inverse relationship between leukocyte TL and hippocampus volume among All subjects gave written informed consent, all clinical investigations were conducted according to the principles expressed in the declaration of Helsinki, and the study was approved by The Regional Ethical Review Board in Umeå, Sweden.APOE ε3/ε3 carriers, 20 ε3/ε4 carriers and 10 were ε4/ε4 carriers. APOE ε4 homozygotes were recruited initially. Following this, age-, gender-, and education-matched ε3/ε3 and ε3/ε4 carriers were recruited. All subjects were non-demented , aged 49 to 79; all recruited from the Betula study. The Betula study is an ongoing prospective cohort study initiated in 1988 aimed at exploring various aspects of memory, health and aging. To date, approximately 4500 participants have been enrolled in the Betula study, all randomly selected from the population registry of Umeå municipality, Sweden. The study population has been validated to conform well with the general population of northern Sweden. Exclusion criteria for enrollment into the Betula project were dementia, mental retardation, serious visual or auditory handicaps, not having Swedish as a mother tongue, or any other feature that would compromise the ability to comply with the study protocol . The ratio of the mean telomere repeat copy number to the mean single copy gene copy number (T/S ratio) as related to that of a reference sample (DNA from the cell line CCRF-CEM) reflects relative TLs. Relative TLs (measured in relative T/S ratios) were converted to telomere restriction fragment lengths (measured in base pairs) based on correlation data derived from samples measured using both quantitative real-time PCR and the conventional southern blot method. Plasma cotinine was measured on the IMMULITE® 2000 Immunoassay System . Cotinine is the major metabolite of nicotine, and measurement of it is the favored method of detecting use of tobacco products Leukocyte DNA was extracted from whole blood using standard procedures. All blood samples were drawn within a 5 year time-frame of the MRI examination. Magnetic resonance imaging was performed using a Philips Intera 1.5 T scanner . Results from structural scans carried out on this sample have been reported previously t-test. Distributions of categorical variables were tested using the Pearson chi-square test. The rate of TL shortening with age was determined using linear regression. To avoid undue influence of outliers, and due to low number of participants in each APOE subgroup, Spearman's rho (non-parametric) was used to evaluate correlations between TL and hippocampus volume, and between age and hippocampus volume. TL was age adjusted, and body height was used to adjust hippocampal volumes for differences in body (and head) size via analysis of covariance Means were compared using the independent-samples


PPARγ is expressed in structural, immune, and inflammatory cells in the lung. PPARγ is involved in mucin production, and PPARγ agonists can inhibit mucin synthesis both in vitro and in vivo. These findings suggest that PPARγ is a novel target in the treatment of AMH and that further work on this transcription factor may lead to new therapies for chronic airway inflammatory diseases.Airway mucus hypersecretion (AMH) is a key pathophysiological feature of chronic airway inflammatory diseases such as bronchial asthma, cystic fibrosis, and chronic obstructive pulmonary disease. AMH contributes to the pathogenesis of chronic airway inflammatory diseases, and it is associated with reduced lung function and high rates of hospitalization and mortality. It has been suggested that AMH should be a target in the treatment of chronic airway inflammatory diseases. Recent evidence suggests that a key regulator of airway inflammation, hyperresponsiveness, and remodeling is peroxisome proliferator-activated receptor gamma (PPAR Airway mucus hypersecretion (AMH) is a common pathological feature in chronic airway inflammatory diseases such as asthma, chronic obstructive pulmonary disease (COPD), and cystic fibrosis (CF). Growing studies have suggested that AMH is associated with the progression of chronic airway inflammatory diseases and it is a significant contributor to morbidity and mortality. To control, AMH plays an important role in the treatment of chronic airway inflammatory diseases; however, effective therapies that target AMH are lacking , 2.Peroxisome proliferator-activated receptors (PPARs) are a family of ligand-activated transcription factors belonging to the nuclear hormone receptor family. PPARs are related to retinoid, glucocorticoid, and thyroid hormone receptors . They reγ is the most extensively studied PPAR subtype. It is involved in a series of lung diseases, including lung fibrosis, pulmonary vascular diseases, acute lung injury, and lung cancer [γ activation reduces the synthesis and release of immunomodulatory cytokines from various cell types that participate in the regulation of inflammatory and immune processes. Specifically, mounting evidence suggests that PPARγ plays important roles in regulating processes related to airway inflammation, airway remodeling, and airway hyperresponsiveness, indicating that PPARγ and its ligand show potential as targets to develop treatments for chronic airway inflammatory diseases [γ has been implicated in AMH. Recent studies have shown that chronic airway inflammatory diseases in humans are associated with altered PPARγ expression and that PPARγ ligand can attenuate AMH in both in vitro and in vivo experimental models. This paper will summarize recent work implicating PPARγ in AMH.PPARg cancer –10. Besidiseases . In factMucus secretion is essential for protecting airways. It is vital for air humidification, warming, and cleaning . HoweverStudies suggest that AMH is not merely a clinical symptom but instead a critical factor in the pathogenesis of chronic airway inflammatory diseases. AMH is associated with greater susceptibility to COPD, a decline in forced expiratory volume in one second, hospitalization, and excess mortality –16. The Pseudomonas aeruginosa and other organisms. AMH contributes to CF morbidity by increasing the frequency and severity of pulmonary infections as well as by impairing lung function [AMH has long been recognized as a major cause of death in asthma. One study found luminal occlusions covering 20–100% of the cross-sectional area in patients who died from asthma, leading the authors to conclude that luminal obstruction of airways by an exudate composed of mucus and cells is a major contributor to asthma fatality . In addifunction . P.  aeruginosa proteases are well-known inflammatory stimuli that can cause significant inflammatory responses, which result in goblet cell metaplasia and hyperplasia, leading in turn to mucin overproduction and mucus hypersecretion in airways [α (TNF-α), interleukin (IL)-1β, IL-6, IL-8, IL-13, and IL-17 can upregulate the expression of MUC5AC, the marker of goblet cell metaplasia, through different signaling pathways [Abnormal inflammatory response is the major component of chronic airway inflammatory diseases. Such inflammatory response has been associated with AMH in airway diseases. Numerous studies have demonstrated that inflammatory stimuli and mediators/cytokines contribute to excess mucin synthesis and mucus secretion in airways. For instance, cigarette smoke, neutrophil elastase, and  airways –24. Inflpathways –29. ThisIn summary, AMH arising as a result of inflammatory processes can lead to physiologically and clinically measurable mechanical airway obstruction in patients with chronic airway inflammatory diseases. In this way, AMH significantly affects the pathogenesis, progression, and prognosis of these diseases. Therapies that aim to control the inflammation that triggers AMH may be effective at treating chronic airway inflammatory diseases –32.γ is expressed in many types of structural cells in the lung, including fibroblasts, ciliated airway epithelial cells, alveolar type II pneumocytes, and airway smooth muscle cells [γ-knockout mice has highlighted the role of this transcription factor in animal models. A specific PPARγ deletion in airway epithelium produced persistent enlargement of the airspaces in adult mice, which showed more severe emphysema symptoms and higher macrophage numbers after exposure to cigarette smoke than did mice carrying the PPARγ  gene [γ is required for proper lung maturation and response to injury. PPARle cells , 4, 32. Rγ  gene . These rγ is also present in several types of inflammatory, immune cells in the lung, where it exerts anti-inflammatory and immuno-modulatory effects. Specially, PPARγ has been found in macrophages, eosinophils, T cells, B cells, and dendritic cells [PPARic cells –38. It pγ expression and function may be associated with chronic airway inflammatory diseases and that stimulation of this receptor may attenuate inflammatory responses. A study examining the expression of PPARγ in lung tissues of patients with COPD found the protein to be present mainly in alveolar epithelial cells and smooth muscle cells in the bronchial walls. The intensity of PPARγ was inversely proportional to the severity of the COPD, with the signal stronger in patients with milder COPD [γ is expressed in CF- and non-CF-type human airway epithelial cells, but appears to be either less abundant, less functional in binding its target DNA sequence, or both, in CF [γ mRNA and protein expression in alveolar macrophages [γ agonists in the treatment of chronic lung inflammatory diseases.Increasing evidence suggests that altered PPARder COPD . Anotherh, in CF . In asthrophages . These fγ and airway inflammatory diseases, several studies have investigated the therapeutic role of the receptor and its ligand in cellular and animal models of these diseases. Administration of PPARγ ligand to BALB/c mice sensitized and challenged with ovalbumin (OVA) reduced the levels of proinflammatory mediators in bronchoalveolar lavage fluid (BALF) and attenuated the inflammatory response in the lung [γ agonist troglitazone blocked the ability of phorbol 12-myristate 13-acetate to stimulate an increase in TNF-α levels. However, this effect of troglitazone required the presence of MUC1/Muc1 [γ exerts an anti-inflammatory effect by stimulating MUC1/Muc1 expression, which then blocks the production of TNF-α/IL-8 induced by phorbol 12-myristate 13-acetate in airway epithelial cells [γ plays an essential role in the pathway through which monocyte/macrophage-derived microparticles activate NF-κB and ultimately induce upregulation of proinflammatory mediators in human lung epithelial cells [Building on the evidence linking PPARthe lung , 42, 43.UC1/Muc1 . The autal cells . In addial cells .γ is thought to be due mainly to its ability to downregulate proinflammatory gene expression. It does so by sequestering shared coactivators, reducing the ability of inflammatory transcription factors to bind their target DNA. The second way is through ligand-dependent transrepression, preventing other transcription factors' association with DNA sequences. At the same time, PPARγ can increase the transcription of anti-inflammatory genes. Agonist binding to PPARγ induces a conformational change in the receptor, allowing the dissociation of corepressors and the association of coactivator molecules. This allows the formation of PPARγ dimmers or heterodimers and subsequent interaction with peroxisome proliferator response elements, which increases anti-inflammatory gene transcription [The anti-inflammatory effect of PPARcription .γ participates in both physiological and pathological processes in chronic airway inflammatory diseases. These studies make a strong case for PPARγ activation as a potential treatment for chronic airway inflammatory diseases. Taken together, these findings suggest that PPARγ and its ligand help regulate airway inflammation, several studies have investigated the potential involvement of PPARγ and its ligand in AMH.Since the inflammatory response is the major contributor to AMH, and PPARγ ligand on mucin production in human airway epithelial NCI-H292 cells exposed to cigarette smoke [α and MUC5AC production, which was attenuated by administration of the PPARγ agonist rosiglitazone. Rosiglitazone increased the expression of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and inhibited the PI3K/Akt pathway. PTEN antagonizes PI3K-mediated signaling, whose role in mucin production is well documented [γ can significantly inhibit cigarette smoke-induced mucin production, raising the possibility that this approach may be useful in the clinic. Indeed, the study authors proposed that using a PPARγ agonist to activate PTEN and inhibit PI3K/Akt may be a potential therapy for cigarette smoke-induced mucin secretory diseases [Lee and colleagues carried out the first study to examine the role of PPARte smoke . Exposurcumented , 48. Thediseases .β, IL-8, and TNF-α) in BALF. Treating the rats with rosiglitazone before acrolein exposure attenuated these changes in a dose-dependent manner [κB activation, which is well known to play an important role in regulating not only multiple inflammatory cytokines but also AMH in inflammatory lung diseases [κB activation by rosiglitazone may explain the observed downregulation of proinflammatory mediators and the reduction in AMH. From these findings, Liu et al. concluded that ligand-induced activation of PPARγ may be a novel therapy for AMH in chronic airway inflammatory diseases such as COPD [Subsequent studies evaluated the effect of rosiglitazone on airway mucin production in animal models of AMH. Exposing rats for two weeks to acrolein, one of the most toxic components in cigarette smoke, induced goblet cell hyperplasia in bronchial epithelium and upregulated MUC5AC mRNA and protein expression in rat lungs. These changes were associated with airway inflammation, as evidenced by the increased numbers of inflammatory cells and levels of inflammatory cytokines  or adenovirus carrying PPARγ2 cDNA showed a lower percentage of airway epithelium staining positive with PAS than did exposed mice that did not receive agonists or adenovirus [γ agonists support protective remodeling of the airway wall by regulating TGF-β or TGF-β1 and collagen deposition. Results from both of these mouse models support the possibility of using PPARγ agonist to treat AMH in humans.AMH is also an important pathologic feature of asthma, which is characterized by nonspecific airway inflammation. Several studies have examined the involvement of PPARtely 75% . In a dienovirus . In addiγ and AMH. Activation of PPARγ by rosiglitazone or pioglitazone markedly reduced mRNA expression of matrix metalloproteinase-9 (MMP-9) and inhibited TNF-α  induced MMP-9 gelatinolytic activity [γ may attenuate AMH through a pathway involving MMP-9. Simvastatin is a statin that modulates inflammatory processes and that can inhibit mucin production and AMH [γ activation [γ. It seems clear from these diverse lines of evidence that the effects of PPARγ and its ligand on AMH are mediated by multiple mechanisms. Further work is needed to identify new ligands of PPARγ and investigate the signaling pathways affected by the receptor. In addition to these observations in cell lines or animal models, several lines of indirect evidence link activation of PPARactivity . Since Mactivity , these r and AMH . A recentivation , which mγ is involved in AMH and that PPARγ ligand suppresses AMH by inhibiting proinflammatory cytokines and inflammatory signaling pathways. Therefore, PPARγ agonists may have potential as treatments for AMH in chronic airway inflammatory diseases.In summary, substantial evidences indicate that PPARγ is involved in AMH and that PPARγ agonists inhibit AMH, there is still a long road ahead before laboratory studies can be translated to the clinic. For one thing, the pathogenesis of chronic airway inflammatory diseases such as COPD, asthma, and CF is extremely complicated. Only a few studies, most of them descriptive, have directly investigated the role of PPARγ and PPARγ ligand in AMH. As a result, the signaling pathways affected by PPARγ or PPARγ agonists remain poorly understood. Further investigation of these areas will be required to define the effects of PPARγ ligands on airway epithelial cells and the mechanisms by which these pro- and/or anti-inflammatory responses occur. Although several studies have suggested that PPARγ in the clinic is that the route of PPARγ agonist administration may be important. In Honda's study [A second consideration for targeting PPAR's study , treatin's study . Oral ga's study , while o's study . Future γ agonists in patients with chronic airway inflammatory disease should be carefully examined. Recent studies suggest an association between rosiglitazone treatment and increased risk of cardiovascular events in patients with type 2 diabetes [γ agonist is safe for patients with chronic airway inflammatory diseases. Finally, the safety of PPARdiabetes , 61. Thuγ and PPARγ ligand as potential target in treating AMH. Corticosteroid is the first-line anti-inflammatory drug used to chronic airway inflammatory diseases, but its therapeutic effects are controversial, and physicians have begun to pay attention to its undesirable side effects [γ is involved in mucin production and that PPARγ agonists inhibit AMH both in vitro and in vivo. Indeed, a recent clinical trial found that rosiglitazone improved lung function in steroid-resistant asthma patients [γ agonist treatment. Thus, PPARγ agonists may represent a novel class of pharmacological agents useful in the management of AMH in chronic airway inflammatory diseases. The present paper describes the clinical importance of AMH in the pathogenesis of chronic airway inflammatory diseases and makes the case for exploring PPAR effects . In fact effects , so therpatients , providi


The probe DNA is immobilized on a self-assembled monolayer, a conducting polymer film, or a layer of nanostructures on the electrode such that desired probe DNA would selectively hybridize with target DNA. The rate of charge transfer from the electrode thus modified to a redox indicator, e.g., [Fe(CN)6]3−/4−, which is measured by EIS in the form of charge transfer resistance (Rct), is modulated by whether or not, as well as how much, the intended target DNA is selectively hybridized. Efforts made to enhance the selectivity as well as the sensitivity of DNA sensors and to reduce the EIS measurement time are briefly described along with brief future perspectives in developing DNA sensors.Recent advances in label free DNA hybridization sensors employing electrochemical impedance spectroscopy (EIS) as a detection tool are reviewed. These sensors are based on the modulation of the blocking ability of an electrode modified with a probe DNA by an analyte,  Thus, the Rct values indicate how crowded the electrode surface is when it is modified by a functional molecule, which is capable of selectively capturing a given analyte. The Rct values are determined by how selective binding has taken place with the analyte and how much analyte is in the test solution.An electrode/electrolyte interface may be simplified as shown by the schematic diagram in Rct, is associated with the energy barrier for electron transfer to/from the redox indicator approaching to the electrode surface, which is determined by the change in the crowdedness of the probe layer caused by its binding with target DNA. In a non-faradaic sensor, the capacitance of the probe layer is a main indicator exhibiting the conformational changes of double-stranded DNA due to its hybridization. Katz and Willner have published an excellent review on various biosensors including immunosensors, enzyme based sensors, and DNA sensors employing faradaic and non-faradaic impedance spectroscopy as a detection tool 3−/4− redox indicator. Later, they examined eight metal ions including alkaline earth metal ions (Mg2+ and Ca2+), trivalent ions (Al3+ and La3+), and divalent transition metal ions  and reported that these metal ions cause decreases in Rct values. The difference of the charge transfer resistance (ΔRct) of ds-DNA films in the presence and absence of these metal ions was different and particular to a given metal ion 3-/4-, leading to detection limits of 1 × 10−12 ∼ 1 × 10−13 M 3−/4− redox indicator during the hybridization is related to the probe density obtained by controlling the fraction of thiolated probe DNA in the mixed solution. As a result, the increase in charge-transfer resistance was observed only above a threshold probe surface density of 2.5 × 1012 cm−2 due to the repulsion between negatively charged DNA and the negative redox indicator ions, i.e., the [Fe(CN)6]3−/4− pair. The optimum probe density was found to be 5.4 × 1012 cm−2, which was obtained from a solution containing both thoiolated DNA and MCH with a DNA mole fraction of 20%. Kjällman et al. employed thiolated polyethylene glycol [PEG: CH3-(CH2CH2O)6-CH2CH2SH] as an alternative for MCH for controlling the probe density and blocking the non-specific binding 3−/4− redox indicator. As described above, the PAMAM dendrimer can be utilized for the immobilization of probe DNA or the signal enhancement via target-labeling. Probe DNA of an appropriate density obtained by a cone-shaped dendrimer showed very high specificity to the complementary target, leading to different responses of Rct values in the presence of [Fe(CN)6]3−/4−depending on the location of the mismatched pair in the duplex  film, which was electrochemically synthesized from the pyrrole monomer and 4-(3-pyrrolyl) butanoic acid, was used as a DNA immobilization platform 3−/4−, obtained from Rct value:vs. Ag/AgCl), amine-modified probe DNA was covalently attached to the carboxyl group of the side chain of the copolymer through the (1-ethyl-3-(3-dimethylaminopropyl) (EDC) carbodiimide coupling. When the hybridization efficiency was compared with the results obtained by faradaic impedance measurements, the poly(Py-co-PDDA) film containing more conjugated side chains showed the largest increase in Rct upon complementary hybridization as shown in As a simple approach, polypyrrole (PPy) was used directly for the immobilization of a DNA probe based on the electrostatic attraction between electrochemically oxidized-PPy and negatively charged oligonucleotides ,45. The PPy film ,47. In tPPy film ,49. Pengplatform ,51. On aid (PPA) ,53. Afteet al. utilized the thiophene matrix covalently coupled with amine modified-probe DNA as a platform for the DNA sensor 3−/4− redox indicator was remarkably increased upon hybridization for the complementary target due to the high density negative charges, space blocking, and semiconducting properties of the CdS nanoparticles, which lowered the detection limit down to 1.43 × 10−10 M. Also, gold nanoparticles deposited on the gold electrode were employed as a platform for the immobilization of thiolated probe DNA [Rct values before and after hybridization showed a linear relation with the concentration of the target in a range of 2.0 × 10−12 ∼ 9.0 × 10−8 M. Bonnani et al. utilized a streptavidin-coated gold nanoparticles for improving the Rct signal [Rct signal. The relative Rct variance for the [Fe(CN)6]3−/4− pair was magnified with an increase in the target concentration due to the electrostatic repulsion and steric hindrances. The detection limit was improved from 26.5 pmol to 11.8 pmol in comparison to the non-amplification method. In their more recent report, they described the immobilization of the amine-terminated probe DNA by carbodiimide chemistry on the screen-printed electrode, which was modified by carboxylated multi-walled carbon nanotubes [Rct values were monitored in a concentration range of thrombin from 0.12 nM to 30 nM. Using the [Fe(CN)6]3−/4− redox indicator, Deng et al. detected thrombin through the increase in Rct on the sandwich system of aptamer/thrombin/ aptamer functionalized Au nanoparticles for a cascaded signal amplification 3−/4− redox indicator 3−/4− indicator after hybridization showing a detection limit of 2.3 × 10−13 M 3−/4− indicator present to probe how far the reaction proceeded 3−/4− redox indicator [Rct value was decreased upon addition of serotonin instead due to a different chemistry taking place from that for the HbA1c sensor. Both cases indicate that monitoring the change in Rct values is much more sensitive than monitoring currents by voltammetric or amperometric methods. We should also point out that the FTEIS technique, although we have not described in detail here, allows one to record a full spectrum in a short period, as short as less than 0.1 s depending on the electrochemical reversibility of the redox indicator used [While we have been describing only the EIS method of detection for the DNA sensors thus far, both amperometric and voltammetric detection methods had widely been used in earlier studies . Althoug(CVs) do ,91. In oroceeded . When thtor used -96.6.We have briefly reviewed recent advances made in DNA sensors employing EIS as a detection tool among many other biosensors using similar platforms -101. ThrDespite the advantages of EIS as a detection tool described earlier in this review, it also suffers from a few disadvantages and limitations. In most cases, its sensitivity still cannot match that of the fluorescence detection as the sensitivity of the latter method is directly related to the intensity of the excitation source. Another limitation arises from the requirement that the EIS detection must be used only at an electrode, while any surfaces may serve as a platform for the fluorescence detection. Again, we described how the surface chemistry can enhance the sensitivity as well as the selectivity.It often takes too long to acquire an EIS spectrum in a reasonably wide frequency range employing the traditional frequency response analyzer. For this reason, a series of relative values of impedances or capacitances at a fixed frequency have been frequently utilized for real-time sensing ,57, whic


Objectives. To apply modern text-mining methods to identify candidate herbs and formulae for the treatment of diabetic nephropathy. Methods. The method we developed includes three steps: (1) identification of candidate ancient terms; (2) systemic search and assessment of medical records written in classical Chinese; (3) preliminary evaluation of the effect and safety of candidates. Results. Ancient terms Xia Xiao, Shen Xiao, and Xiao Shen were determined as the most likely to correspond with diabetic nephropathy and used in text mining. A total of 80 Chinese formulae for treating conditions congruent with diabetic nephropathy recorded in medical books from Tang Dynasty to Qing Dynasty were collected. Sao si tang  was chosen to show the process of preliminary evaluation of the candidates. It had promising potential for development as new agent for the treatment of diabetic nephropathy. However, further investigations about the safety to patients with renal insufficiency are still needed. Conclusions. The methods developed in this study offer a targeted approach to identifying traditional herbs and/or formulae as candidates for further investigation in the search for new drugs for modern disease. However, more effort is still required to improve our techniques, especially with regard to compound formulae. Natural products used in traditional medicine have historically been invaluable for drug development , 2. SuccHowever, as the years passed, diseases and their names changed, leading to the disassociation between the traditional and modern medical terminologies. Given the voluminous content of the traditional Chinese medical literature, conducting searches to identify potential drug candidates is challenging. Additionally, the effects of classical formulae for the treatment of modern diseases still need to be assessed. All of these aspects present obstacles to the effective and efficient use of the classical literature resources for therapeutic product discovery. Consequently, modern approaches that can mine these classical medical records of traditional Chinese medicine need to develop. Over the last five years, through the International Research Network for Traditional and Complementary Medicine (IRN-TCM), we have developed and refined methods for text mining of the traditional Chinese medicine classical literature to identify candidate herbs and herbal combinations that show potential for further research –13.Diabetic nephropathy is the most common cause of end-stage renal disease around the world and is characterized by rapid progression and a poor prognosis . With thThe project involved three parts: (1) identification of classical terms that could refer to diabetic nephropathy; (2) text mining of the classical Chinese medical literature; and (3) preliminary evaluation of the effect and safety of candidates on diabetic nephropathy and the selection of candidates for further drug discovery efforts.In order to identify all the classical terms that could have referred to diabetic nephropathy, literature searches were conducted. Articles that focused on original researches related to classical medical terms and on the experience of venerable TCM doctors were retrieved from the Chinese databases CNKI, VIP, Wan Fang, CBM, and TCM online. Medical textbooks for undergraduate and postgraduate teaching issued by the state and medical monographs on diabetic nephropathy were also collected through the library of Guangzhou University of Chinese Medicine.Two authors extracted the classical terms related to diabetic nephropathy that were mentioned in these sources and calculated the frequency of mention for each term. In order to obtain expert opinion on which terms were more corresponding with diabetic nephropathy, a questionnaire was designed and distributed to traditional medicine hospitals around China. Heads of the nephrology department in these hospitals who had more than 10 years of clinical experience in classical medical Chinese were consulted.The consulting questionnaire included all the classical terms, classical medical records describing their clinical manifestations, and the clinical features of diabetic nephropathy according to the diagnostic criteria of modern medicine. Experts were required to gauge the degree of consistency between the classical term and the modern conception of diabetic nephropathy by comparing their clinical manifestations descriptions. Frequencies of each classical term mentioned in research articles, empirical articles, textbooks, and medical monographs were attached as a reference.The degree of consistency was divided into five categories: completely consistent (5 points), mostly consistent (4 points), partly consistent (3 points), seldom consistent (2 points), and completely inconsistent (1 point). Experts had to tick only one category for each classical term. Total score of each classical term was calculated by adding the points experts ticked. Scoring rate of each classical term was full score divided by its total score and then multiplied by 100%. Full score was 5 points multiplied by the number of returned questionnaires.These classical terms with scoring rate more than 50 percent were regarded as identified terms by expert consultation for further verification. Their corresponding modern diseases were retrieved in the textbooks and monographs of Chinese Internal Medicine, monographs of kidney disease of Chinese Medicine, and dictionaries of Chinese Medicine via the library of Guangzhou University of Chinese Medicine. The mentioned frequencies of each modern disease were counted.Classical terms which have corresponding modern diseases not limited to diabetic nephropathy or targeting many organs not mainly in kidney were excluded. Classical terms with corresponding modern diseases which refer to kidney damages occurring in diabetes mellitus were included and used in ancient literature searching.“Encyclopedia of Traditional Chinese Medicine” , which includes 1009 different Chinese medical books written before the emergence of the People's Republic of China (1949 AD) , was selThe information about the treatments of these included classical terms was extracted, including the titles and completion dates of the books, all records related to therapies for disorders congruent with diabetic nephropathy, and the formulae used for treating these disorders. Ancient formulae targeting incongruent disorders with diabetic nephropathy confirmed by two authors, respectively, were excluded. Discrepancies were resolved by a third author, who made the final decision. The frequency of citation of each included formula was calculated. Formulae with higher recorded frequency were selected as candidates for further work in drug discovery for diabetic nephropathy.A preliminary evaluation of the effect of candidates on diabetic nephropathy was conducted by searching the databases PubMed (January 1966 to June 2012), EMBASE (January 1985 to June 2012), the Cochrane Library, and clinicalTrials.gov to locate studies on the clinical application and experimental research on candidate formulae and their components.Database searches resulted in the inclusion of 91 original research articles and 25 articles on the empirical experience of venerable TCM doctors that mentioned classical terms for DN. 60 medical monographs and 11 textbooks that included sections on diabetic nephropathy were selected via the library of Guangzhou University of Chinese Medicine.A total of 31 classical terms associated with diabetic nephropathy were collected for expert consultation . Frequen水肿), Shen Xiao (肾消), Niao Zhuo (尿浊), Guan Ge (关格), Xu Lao (虚劳), Xia Xiao (下消), Xiao Ke (消渴), and Xiao Shen (消肾) were more than 50 percent . Scoring rates of Shui Zhong  include renal edema, cardiac edema, nutritional edema, endocrine edema, hepatic edema, and edema of unknown reason. Besides diabetic nephropathy, renal edema also refers to acute or chronic glomerulonephritis, nephrotic syndrome, other secondary glomerular diseases (such as lupus nephritis), and chronic renal failure. Xu Lao (虚劳) is considered as chronic and consumptive disease involving multisystems and multiorgans, especially organ function decline or failure. Guan Ge (关格) is regarded as chronic renal failure, acute renal failure, uremia period, ileus, and esophageal carcinoma. Niao Zhuo (尿浊) refers to chyluria, phosphaturia, filariasis, urinary system infection, urinary system cancer, tuberculosis, and so on. Xiao Ke (消渴) mainly refers to diabetes mellitus (Corresponding modern diseases of Shui Zhong (mellitus .肾消), Xia Xiao (下消), and Xiao Shen (消肾) were not regarded as independent diseases in textbooks and monographs of Chinese Internal Medicine, and monographs of kidney disease of Chinese Medicine. They were mentioned in Xiao Ke (消渴) when kidney damage occurs (Shen Xiao (e occurs .消肾) the Bei Ji Qian Jin Yao Fang, written by Sun Si-miao during the Tang Dynasty (652 AD), provides the following description: “消肾) diseasePatients with symptoms such as fever due to deficiency, thirst but not drinking more water, frequent urination, turbid urine and thready pulse were often diagnosed as Xiao Shen , the following definition is provided: “下消) with the symptoms of dark urine, turbid urine, gloomy complexion, muscle wasting, is also called Shen Xiao (肾消), as the disease location is in the kidney Xia Xiao , provided the following linkage with diabetes: “肾消) progresses from Xiao Ke (消渴), with the symptoms of polydipsia, polyuria and turbid urineShen Xiao (.”The following three extracts are examples of descriptions consistent with DN . In rela下消) refers to Shen Xiao (肾消) and Xiao Shen (消肾).The relationship between the three ancient terms was described in 8 dictionaries of Chinese Medicine. Xia Xiao (消肾) refers to Xia Xiao (下消) and Shen Xiao (肾消). But besides Xia Xiao (下消), Shen Xiao (肾消) also refers to Qiang Zhong (强中), which is called priapism in modern times. Therefore, Xia Xiao (下消), Shen Xiao (肾消), and Xiao Shen (消肾) were considered more corresponding with diabetic nephropathy. However, ancient records about the symptoms of priapism should be excluded during the ancient formulae information extraction.Xiao Shen (下消), Shen Xiao (肾消), and Xiao Shen (消肾) via “Encyclopedia of Traditional Chinese Medicine.” Ancient records which were thought to be corresponding with the symptoms of priapism were not included for formulae extraction.This study searched ancient records of Xia Xiao (黄芪饮), Xuan bu wan (宣补丸), and E jiao tang (阿胶汤). Eighteen formulae were recorded more than 5 times. The top eight formulae were, in the order, Liu wei di huang wan (六味地黄丸), Jia jian shen qi wan (加味肾气丸), Bai fu ling wan (白茯苓丸), Si wu tang (四物汤), Sao si tang (缫丝汤), Hui xiang san (茴香散), Lu rong wan (鹿茸丸), and Ren shen san (人参散)  to Qing Dynasty (1644 AD to 1912 AD). The earliest formulae for treating diabetic nephropathy recorded in Tang dynasty were Huang qi yin (an (人参散) .缫丝汤), Hui xiang san (茴香散), Si wu tang (四物汤), Ge gen wan fang (葛根丸), and Gu ben wan (固本丸). Liu wei di huang wan (六味地黄丸), Jia jian shen qi wan (加减肾气丸), Lu rong wan (鹿茸丸), Hu fen san (胡粉散), and E jiao tang (阿胶汤) contain more than 5 and fewer than 10 ingredients. Bai fu ling wan (白茯苓丸), Shen li san (肾沥散), Xuan bu wan fang (宣补丸方), Cong rong wan (苁蓉丸), Shuang bu wan (双补丸), Gou qi zi wan (枸杞子丸), and Ping bu wan (平补丸) contained more than 10 ingredients  ” ranked fifth and was the simplest since it only contained one ingredient—silkworm and/or silk cocoon.After identification of the candidate formulae, preliminary evaluation of their effect on diabetic nephropathy was undertaken. This began with the simple, high frequency formulae. Among the 18 formulae, “Sao si tang (Yi Xue Zheng Zhuan written by Yu Tuan during the Ming Dynasty (1515 AD). In reference to the inherited formula Reeling Silk Decoction, he wrote that “it has an excellent effect on Shen Xiao with the symptoms of turbid urine, polydipsia and excessive appetite but the person loses weight….. the effect of the hot water used in reeling silk (i.e. Reeling Silk Decoction) is best. If this is not available, it can be replaced by a decoction of silkworm cocoon or silk floss.”—from Yi Xue Zheng Zhuan [The earliest record of its use was in ng Zhuan .Based on this report and subsequent repeated citation of this remedy by other authors, we conducted the literature search of the modern studies regarding the silkworm, its related products, and its active ingredients, for treating diabetic nephropathy in order to investigate whether this simple formula could have the potential to be developed into a new agent for diabetic nephropathy.No studies of Reeling Silk Decoction were located, but there have been considerable studies involving silkworm, its related products, and its active ingredients. 202 articles describing the active ingredients of the silkworm and its products for diabetic nephropathy were retrieved in a search of the modern literature .According to modern studies, the silkworm and its products are rich in various active substances such as alkaloids, flavanoids, and silk protein hydrolysates.1-Deoxynojirimycin (DNJ) is a major component of the alkaloids in silkworm . A cliniβ1 (TGF-β1) expression [κB (NF-κB) expression, which may be involved in the pathogenesis of proteinuria in diabetic nephropathy [Among the flavanoids, which have been purified and identified from the sericin layer of silkworm cocoons , quercetpression  and amelpression , which hpression . Additiohropathy , 29.Additionally, the concentrations of 1-DNJ and the activities of quercetin in silkworm are higher than in mulberry leaves, which are the only food source of silkworm, because of the biotransformation in the silkworm body –35.Therefore, components of Reeling Silk Decoction have demonstrated promising potential for development as new agents for the treatment of diabetic nephropathy. However, its safety for patients with renal insufficiency should be evaluated in further investigations.The methods used in classical traditional Chinese medicine, which have been “clinically” tested for thousands of years, continue to play an indispensable role in the treatment of chronic diseases in Asian countries. It has also become an important source of drug discovery for Western scholars and pharmacologists. However, barriers such as the disassociation between the traditional and modern medical terminologies, and the voluminous content of traditional Chinese medical literature, have slowed the pace of drug discoverer using the resources of the classical literature. The use of modern technology and methods for text mining of the traditional Chinese medicine classical literature can provide an approach to accelerating this process.According to the method we developed, the process of drug discovery from the classical medical literature includes three main steps: (1) identification of candidate classical terms; (2) systemic search and analysis of classical medical records; (3) preliminary evaluation of the effects and safety of the candidates.The usual method for identifying ancient terms corresponding with modern disease is based mainly on narrative reviews of the classical literature. However, the result in this study indicated that correspondence between ancient terms and modern disease was overlapping, rather than there being a one-to-one correspondence. This phenomenon also appeared in age-related dementia and memory impairment . So the 水肿) was named after symptom of a visible edema caused by discords in many systems. Besides diabetic nephropathy, chronic or acute glomerulonephritis, nephrotic syndrome, and other secondary glomerular diseases may result in renal edema as well, which is usually characterized by facial or lower limb swelling due to water-sodium retention or hypoproteinemia. Chronic renal failure was one of the modern diseases corresponding with Guan Ge (关格) and Xu Lao (虚劳). It was the serious end stage of all the progressed chronic kidney diseases, not only diabetic nephropathy. Xiao Ke (消渴) was regarded as diabetes mellitus, which referred more to diabetes without kidney damage. And the modern diseases of Niao Zhuo (尿浊) would prefer chyluria, tuberculosis, urinary system infection, and cancer, rather than diabetic nephropathy. Therefore, it was difficult to identify that if the ancient literature describing these classical terms referred to diabetic nephropathy or not. It deserved further researches specifically identifying treatment related to diabetic nephropathy in their ancient records for each of them. Xia Xiao (下消), Shen Xiao (肾消), and Xiao Shen (消肾) which meant kidney damage occurring in diabetes were considered more corresponding with diabetic nephropathy and used in ancient literature text mining. Because Shen Xiao (肾消) also referred to Qiang Zhong (强中), which meant priapism in modern times. Formulae targeted Qiang Zhong (强中) was excluded when formulae extracting.Among these identified ancient terms, Shui Zhong (The two-way confirmation of terminology correspondence showed the overlap between ancient terms and modern disease more clearly. It was helpful for consistency evaluation between classical text that described these ancient terms and diabetic nephropathy in ancient text mining. However, it would be more convincing if expert consultation was included in modern diseases retrieval, just as done in the classical terms identification of diabetic nephropathy.The systematic search of full texts of medical book firstly required the identification of a suitable collection. Our previous work located fourteen collections of traditional Chinese medical literature that could be used as resources for systematic searches . The mosSince reports about the nephrotoxicity of Chinese Medicine appeared in 1994, and a condition named “Chinese herbs nephropathy”  receivedWe chose Reeling Silk Decoction, which contains only a single agent, as an example in this study, since most researchers pay more attention to individual agents than to compound formulae. This is because a single agent is simple and its effect on modern disease is easier to be elucidated using current technology. However, formulae consisting of only a single agent are not typical of the prescription used in ancient China. In fact, the compound formula containing multiple agents with different roles in treating the diseases is the essence and characteristic feature of traditional Chinese medicine . In our This convergence of the results of text mining of the classical literature and searches of modern biomedical databases illustrates the value of this text-based approach to the selection of candidates for drug discovery endeavours. The use of modern technology for text mining the classical literature of traditional Chinese medicine shows potential and could be an important step towards a brighter future for drug discovery. The methods developed in this study offer a targeted approach to identifying traditional herbs and/or formulae as candidates for further investigation in the search for new drugs for modern diseases. However, more effort is still required to improve our techniques, especially with regard to compound formulae.


Safety analysis has been done for thick-walled circular cylinder under internal and external pressure using transition theory which is based on the concept of generalized principal Lebesgue strain measure. Results have been analyzed theoretically and discussed numerically. From the analysis, it can be concluded that circular cylinder made of functionally graded material is on the safer side of the design as compared to homogeneous cylinder with internal and external pressure, which leads to the idea of “stress saving” that minimizes the possibility of fracture of cylinder. The constantly increasing industrial demand for axisymmetrical cylindrical and spherical components or elements of them has concentrated the attention of designers and scientists on this particular area of activity. The research on the prediction of stresses in thick-walled hollow circular cylinder has never ceased because of the importance of these basic structures in numerous mechanical, civil, electrical, and computer engineering applications. These days in nuclear industry, cylinders subjected to internal and external pressure have become a point of interest due to their application to advanced small and medium-sized light water reactors. For example, steam generator tubes, in which primary coolant flows outside the tubes while secondary water flows inside the tubes are typical examples of cylinders under internal and external pressure. Another example is pipelines under seawater to transport gas, oil, and so forth. Now for design and integrity evaluation of a cylinder under internal and external pressure, one should carefully consider the failure characteristics of a cylinder under internal and external pressure. The failure mechanisms of such type of cylinder might be quite different from those of a typical one under internal pressure. Upon the estimation of load carrying capacity of these thick-walled cylinders under internal and external pressure and combined loading, many numerical and experimental works have been also made to propose relevant design criteria of thick-walled cylinders subjected to internal and external pressure. Plane strain and plane stress analytical solutions of thick hollow cylinder problems in the elastic stress state have been available for many years in standard and advanced textbooks –4. Thickεii by taking the Lebesgue integral of the weighted function εii = ∫0eiiA[(1−2eiiA)]n/2)−1(deiiA = (1/n)[1 − (1−2eiiA)n/2)−1 are constants.In this paper, our aim is to calculate safety factor for thick-walled cylinder made of functionally graded material under internal and external pressure using Seth's transition theory , 14–16. Results obtained have been discussed numerically and depicted graphically. a and b, respectively, subjected to internal pressure p1 and external pressure p2. The non-homogeneity in the cylinder is due to variable compressibility C = C0rk−. The cylinder is taken so large that plane transverse sections remain plane during the expansion, and hence the longitudinal strain is the same for all elements at each stage of the expansion. In cylindrical polar coordinates the displacements are given by [β is a function of d is a constant.Consider a nonhomogeneous thick-walled circular cylinder of internal and external radii given by –16(2)u=n is the measure and β′ = dβ/dr. The generalized strains areI1 = ekk; Tij,  eij are stress and strain tensors, respectively, and δij is Kronecker's delta.For isotropic material, the stress-strain relation in elastic region is given byβ has been obtained by substituting .Equations of equilibrium are all satisfied excepttituting  in 5), , (5)ddr = −∫(C/r)dr.As elastic state can go to plastic state under external loading through a transition state and we consider only the principal stresses. Therefore the transition can take place either through the principal stresses nction R –16 in te[Trr asR=Trr−λnk, we getddrlogR=Trr − Tθθ| is maximum at r = (e2bk)k1/ = r1; that is, yielding starts at r = r1; therefore,By using  in 9) a a9) and Trr=A[exp2.Using , we getquations  yields(r−k)ezz.Also,  TθC0 → 0), (For full plasticity (0 → 0),  becomesParticular Case: Nonhomogeneous Cylinder under Internal Pressure Only. The stresses in fully plastic state arey Sharma .R0 = 0.1 (0.1) 0.5. For a homogeneous (k = 0) circular cylinder, yielding starts at internal surface whereas for a circular cylinder made of nonhomogeneous material , yielding takes place at any radius r where a < r < b depending upon values of C0 and k. Effective pressure is maximum at internal surface for cylinder made of nonhomogeneous as well as homogeneous material. It is seen from To observe the combined effect of pressure on a cylinder made of homogeneous and nonhomogeneous material, graphs have been drawn between pressure and radii ratios It has also been observed from From Figures From the above analysis we can conclude that nonhomogeneous cylinder with internal and external pressure is on the safer side of the design as compared to homogeneous cylinder because nonhomogeneous cylinder requires high pressure for initial yielding as compared to homogeneous cylinder. It has also been concluded that highly compressible nonhomogeneous cylinder is on the safer side of the design as compared to less compressible nonhomogeneous circular cylinder because highly compressible cylinder required high pressure for initial yielding as compared to less compressible nonhomogeneous cylinder, which leads to the idea of “stress saving” that minimizes the possibility of fracture of cylinder.


We have investigated the influence of conserved non-coding sequences (CNSs), DNA patterns retained after an ancient whole genome duplication event, on the breadth of gene expression and the rates of mRNA decay in Arabidopsis thaliana. The absence of CNSs near α duplicate genes was associated with a decrease in breadth of gene expression and slower mRNA decay rates while the presence CNSs near α duplicates was associated with an increase in breadth of gene expression and faster mRNA decay rates. The observed difference in mRNA decay rate was fastest in genes with CNSs in both non-transcribed and transcribed regions, albeit through an unknown mechanism. This study supports the notion that some Arabidopsis CNSs regulate the steady-state mRNA levels through post-transcriptional control mechanisms and that CNSs also play a role in controlling the breadth of gene expression.Steady-state mRNA levels are tightly regulated through a combination of transcriptional and post-transcriptional control mechanisms. The discovery of  Duplication of genetic material has been proposed to be one of the primary evolutionary factors driving organism complexity and occurs at various scales ranging from single gene transpositions to whole genome duplication (WGD) events , and relative levels of gene expression  exhibit higher average levels of expression than other genes in Arabidopsis annotation . Notably, the absence of an annotated 5′-UTR or 3′-UTR was also sufficient to decrease mRNA stability . With the objective of identifying changes in mRNA that could be attributed to CNS presence, we therefore limited our analyses to the 9,958 genes measured by Narsai et al. that contained annotated 5′-UTR, intron, and 3′-UTR sequences. The list of 9,958 genes was separated into the three categories based on gene duplication status: α duplicates, singletons, and non-α duplicates. We considered a p-value ≤ 0.001 significant for all comparisons.Conserved non-coding sequences have been identified in all subgene positions relative to α duplicates [5′-upstream, 5′-UTR, intron, 3′-UTR, and 3′-downstream  had an increased mRNA half-life relative to all genes  = 2.05 × 10−5). Notably, genes with the fastest rates of mRNA decay were also depleted in CNS negative α duplicates relative to the genomic background . Genes with the slowest rates of mRNA decay were enriched in CNS negative α duplicates relative to background . Genes with the slowest rates of mRNA decay had no change in the proportion of CNS positive α duplicates relative to background .These results associate CNS annotation with an increase (CNS positive α duplicates) or decrease (CNS negative α duplicates) in rate of mRNA decay relative to genomic background. In order to verify this trend using a reverse approach we isolated the genes with the fastest rates of mRNA decay  and genes with the slowest rates of mRNA decay  and looked for enrichment or depletion of CNS annotation  could be explained by the combination of transcriptional rate and mRNA decay. Since we observed significant differences in mRNA half-life between CNS positive α duplicates and CNS negative α duplicates, we therefore hypothesized that any variance of gene expression across the microarray datasets could be partially regulated by CNSs through an mRNA decay mechanism. To determine if the observed changes in mRNA decay based on CNS annotation could be attributed to broad (many tissues or conditions) or narrow (few tissues or conditions) gene expression, we examined the sample variance of expression intensity for all genes across the 7,016 expression datasets. We selected the metric τ to quantify the sample variance, as it is similar to the coefficient of variation (CV), but has been reported to be superior compared to CV for measuring breadth of gene expression  for α duplicates with only non-transcribed CNSs relative to all genes  or α duplicates with only non-transcribed CNSs . This suggests that the presence of even a single non-transcribed CNS may be sufficient to reduce mRNA half-life. We attempted to narrow the effect of CNSs on mRNA half-life to individual subgene positions , but were unable to detect any significant differences (data not shown).Within this study we found the presence of CNSs was sufficient to significantly reduce mRNA half-life by ∼0.5 h relative to all genes and ∼1.5 h relative to CNS negative α duplicates  motif that would be bypassed by the way CNSs were discovered.The association of non-transcribed CNSs (5′-upstream and 3′-downstream) with an increased rate of mRNA decay is a surprising finding given that any RNA decay motifs encoded in the CNS would not be present in the preprocessed or mature RNA transcript. The mechanism by which non-transcribed CNSs are influencing the rate of mRNA decay is unknown, but non-transcribed CNSs are in phase with increased mRNA decay. It may be that α duplicates with non-transcribed CNS are associated with motifs that are not encoded within the CNS. For example, a number of genes in cis-regulatory elements responsible for controlling breadth of gene expression. However, only α duplicates with both non-transcribed and transcribed CNSs had lower mRNA half-lives than CNS negative α duplicates, suggesting that the changes in breadth expression are only partially regulated at the level of mRNA decay. The differences in breadth of expression between the gene subsets we tested were also maintained using CV as our metric of breadth of gene expression, although the statistical differences were less defined than τ . The similarity between metrics was due, in part, to a correlation between CV and τ  than CNS negative α duplicates, which suggests that non-transcribed CNSs may contain Arabidopsis . The small differences in AEI were also reflected in mRNA half-life as we found no significant differences in mRNA half-life between α duplicates, singletons and non-α duplicates relative to all genes , further supporting a link between AEI and mRNA half-life. While there was no effect of gene duplication status on mRNA half-life, we did observe a significant decrease in τ for singletons relative to all genes  relative to other genes in s Figure A. Intrigs Figure B. This hcis-regulatory motifs. There is evidence to dispute the third hypothesis, as CNS’ elements have been found to overlap with known gene regulatory networks , it was found that there was still a significant difference in mRNA decay between CNS’ positive α duplicates relative to CNS’ negative α duplicates Table . HoweverRates of mRNA decay have been correlated with several functional classes of genes, such as kinases, plasma membrane proteins and transcription factors  in the TAIR10 genome were considered singletons. There were 5,108 genes that met this criterion in the TAIR10 genome. Any gene that was not an α duplicate or singleton was assigned to the category of non-α duplicates.The list of α duplicates gene pairs were collected from . RMA normalization  represents the maximum expression intensity for the given probe set across all microarray experiments. Genes with a τ = 0 represent expression across all microarrays, while genes expressed in only one microarray will approach τ = 1. Breadth of gene expression was also measured using the coefficient of variation (CV = σ/μ) for each probe set.p ≤ 0.001 were considered significantly enriched. All GOα Duplicates were separated into CNS positive α duplicates, CNS negative α duplicates, α duplicates with only non-transcribed CNSs, α duplicates with only transcribed CNSs, and α duplicates with both non-transcribed and transcribed CNSs. These gene lists were then tested for enrichment of functional terms using a DAVID-like .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.http://www.frontiersin.org/Plant_Genetics_and_Genomics/10.3389/fpls.2013.00129/abstractThe Supplementary Material for this article can be found online at Supplementary Table S1p-values for CNS Gene SubsetsKS Test .Click here for additional data file.Supplementary Table S2p-values for CNS Gene SubsetsWilcoxon Ranked Sum .Click here for additional data file.Supplementary Table S3p-values for CNS’ Gene SubsetsKS Test .Click here for additional data file.Supplementary Table S4p-values for CNS’ Gene SubsetsWilcoxon Ranked Sum .Click here for additional data file.Supplementary Table S5p-value ≤ 0.001)Enriched Function Annotation in Alpha Duplicate Gene Lists based on CNS Annotation (Bonferroni .Click here for additional data file.Supplementary Table S6Arabidopsis Gene mRNA Half-Life and CNS Status.Click here for additional data file.Supplementary Table S7GEO Datasets Used in This Study.Click here for additional data file.