Docosahexaenoic acid (DHA) promotes recovery from postoperative ileus and the repair of the injured intestinal barrier through mast cell-nerve crosstalk.

The objective of this study was to investigate the neuroimmune mechanisms implicated in the enhancement of gastrointestinal function through the administration of oral DHA. Mast cell-deficient mice (KitW-sh) and C57BL/6 mice were used to establish postoperative ileus (POI) models. To further validate our findings, we conducted noncontact coculture experiments involving dorsal root ganglion (DRG) cells, bone marrow-derived mast cells (BMMCs) and T84 cells. Furthermore, the results obtained from investigations conducted on animals and cells were subsequently validated through clinical trials. The administration of oral DHA had ameliorative effects on intestinal barrier injury and postoperative ileus. In a mechanistic manner, the anti-inflammatory effect of DHA was achieved through the activation of transient receptor potential ankyrin 1 (TRPA1) on DRG cells, resulting in the stabilization of mast cells and increasing interleukin 10 (IL-10) secretion in mast cells. Furthermore, the activation of the pro-repair WNT1-inducible signaling protein 1 (WISP-1) signaling pathways by mast cell-derived IL-10 resulted in an enhancement of the intestinal barrier integrity. The current study demonstrated that the neuroimmune interaction between mast cells and nerves played a crucial role in the process of oral DHA improving the intestinal barrier integrity of POI, which further triggered the activation of CREB/WISP-1 signaling in intestinal mucosal cells.

Low-dose Rivaroxaban: is it a safe alternative in Chinese older patients?

PURPOSE: 10 mg rivaroxaban is widely used in the Chinese mainland. This study aims to explore the association between 10 mg once daily rivaroxaban and all-cause mortality in patients with nonvalvular atrial fibrillation (NVAF). METHODS: This observational study enrolled 1131 NVAF patients at the cardiovascular department of the First Affiliated Hospital of Xi'an Jiaotong University. One-year outcomes included all-cause mortality and bleeding were recorded. Cox proportional hazards models and Kaplan-Meier analysis were utilized in the study. RESULTS: In total, 1131 patients (402 no anticoagulants, and 729 rivaroxaban) were included. Cox proportional hazard analysis demonstrated that low-dose rivaroxaban (10 mg, HR: 0.14, 95% CI:(0.07-0.28), P<0.001; 15 mg, HR: 0.20, 95% CI:(0.09-0.43), P<0.001; 20 mg, HR: 0.22, 95% CI:(0.05-0.96), P = 0.044) exhibited lower mortality risk compared to untreated patients. CONCLUSIONS: 10 mg once daily rivaroxaban may provide survival benefits for elderly patients with NVAF.

Associations of blood pressure trajectories in early life with target organ damage in midlife : a 30-year cohort study.

Hypertension is a pivotal factor in cardiovascular risk. However, the association of longitudinal blood pressure (BP) trajectories in the early life and cardiovascular risk assessed by target organ damage (TOD) in adulthood is poorly reported. The objective of this study was to identify the association between systolic BP, diastolic BP, and mean atrial pressure (MAP) trajectories early in life with a single or multiple TOD in later life. We identified BP trajectories from 6 to 45 year-old using group-based trajectory models among 2430 individuals in the Hanzhong Adolescent Hypertension Study and examined the relationship between BP trajectories and cardiovascular risk in later life. Four discrete long-term systolic BP, diastolic BP, and MAP trajectories were identified, namely, low stable, moderate stable, high stable (low increasing), and moderate increasing groups, based on the BP levels at baseline and in the 30-year follow-up. The carotid intima-media thickness were higher in persistently high or increasing trajectories in comparison to the low stable group. Individuals with deteriorative trajectories during early life were at an increased risk of suffering from a single TOD, including left ventricular hypertrophy (LVH) and carotid atherosclerosis (CA) in middle age (36-49 years old). Moreover, higher BP trajectories were correlated with the presence of combined TODs load stage which were assessed by CA, LVH, arteriosclerosis and subclinical renal damage (SRD). Higher longitudinal BP trajectories early in life were associated with increased cardiovascular risk in midlife, and identifying BP trajectories in early life can help screen individuals with TOD later. LVH, left Ventricular Hypertrophy; CA, carotid atherosclerosis; SRD, subclinical renal damage; TOD, target organ damage.

Association of grip strength and comorbidities with all-cause mortality in the older hypertensive adults.

Background: With growing concerns about global population aging, comorbidity, and disability have emerged as key variables that influence the health of the older adults in terms of disease and function. This study sought to examine the impact of comorbidity and impairment using disease and functional status indicators of all-cause mortality in the older adults. Hypertension, which was chosen as the indicator chosen for disease, has the greatest prevalence in the older population. A total of 15 self-reported chronic conditions were added as indicators of comorbidity, and grip strength was chosen as a measure of functional status. The study also evaluated the association between grip strength and comorbidity, as well as its consequences on all-cause death and survival in a hypertensive senior population. Methods: We chose a total of 2,990 hypertensive participants aged ≥60 years whose data for grip strength were collected in the National Health and Nutrition Examination Survey conducted between 2011 and 2014. The association of all-cause death with grip strength and comorbidity was examined using a Cox proportional hazard regression model. The interaction between comorbidity and all-cause mortality, as well as its association with grip strength, was also examined. Results: The hazard ratio [95% confidence intervals (CIs)] for all-cause mortality in the highest grip strength tertile was 0.266 (0.168-0.419), compared to the lowest grip strength tertile. The all-cause mortality decreased with an increase in the number of co-morbidities [2.677 (1.557-4.603) in the group with ≥3 chronic diseases]. The weighted generalized model revealed a negative correlation between grip strength and comorbidities in more than three groups after accounting for all possible variables (ß = -2.219, -3.178 ~ -1.260, p < 0.001). The risk of mortality reduced with increasing grip strength in patients with ≥3 comorbidities (p-value for trend <0.05), but no meaningful difference was found in the interaction between comorbidities and grip strength (p-value for interaction >0.05). Conclusion: In older hypertension patients, grip strength and comorbidities were correlated with all-cause death, and there was a negative correlation between grip strength and comorbidities. Higher grip strength was associated with fewer fatalities in patients with ≥3 comorbidities, suggesting that functional exercise can improve the prognosis of comorbidities.

Association of rate pressure product trajectories at an early age with left ventricular hypertrophy in midlife: a prospective cohort study.

The joint effect of blood pressure (BP) and heart rate (HR) on cardiovascular disease is unclear. Rate pressure product (RPP), the product of systolic BP and HR, is assessed in this study. This study aimed to determine the longitudinal patterns of RPP from childhood to adulthood and to explore the relationship between RPP trajectories in early life and left ventricular hypertrophy (LVH) in midlife. We included individuals with 3 or more RPP values from 7 visits over a 30-year follow-up period in the Hanzhong Adolescent Hypertension Study cohort to fit trajectory groups and performed logistic regression to evaluate the relative risk of developing LVH. Three discrete trajectories in RPP were identified among 2412 participants assessed from childhood to middle-aged adulthood, which were tagged as "low stable," "moderate stable," and "moderate increasing". A higher waist-to-hip ratio, smoking, alcohol consumption, hypertension, diabetes, and hyperlipidemia were associated with increased RPP trajectories. The Cornell voltage product was positively correlated with RPP in 2017 and was higher in the moderate-stable and moderate-increasing groups than in the low-stable group in RPP trajectories. Compared with the low-stable group, the ORs of LVH were 1.65 (1.13, 2.92) for the moderate-stable and 3.56 (2.26, 5.44) for the moderate-increasing group. Subjects with moderate-stable and moderate-increasing trajectories showed higher probabilities of LVH at an elderly age than those in the low stable trajectory group even after adjusting for multiple cardiovascular risk factors. RPP trajectories are identifiable from childhood and are associated with LVH in midlife. Monitoring RPP trajectories from early life may be an effective approach to predict cardiovascular health status later in life.

Analysis of the use of antidepressants in patients from non-psychiatric departments in general hospital.

This study aimed to analyze the use of antidepressants in non-psychiatric departments. The data of patients treated with antidepressants in non-psychiatric departments of the First Affiliated Hospital of Xi 'an Jiaotong University, were collected from 2014 to 2018. The average annual growth rate of antidepressants use was 22.83%. According to the number of patients at discharge, the classification descending order was: SSRIs, Flupenthixol-TCA, SNRIs, TCAs, SARIs, and NaSSA. Sertraline and flupenthixol-melitracen were the main drugs used in SSRIs and Flupenthixol-TCA, respectively. Neurology, Cardiology, and Geriatrics were the main departments prescribing SSRIs, Flupenthixol-TCA and SNRIs. The antidepressants used by all patients were mainly SSRIs according to drug classification, but the specific drug use in unclear diagnosis group was dominated by flupenthixol-melitracen, while it was sertraline in clear diagnosis group. The proportion of Flupenthixol-TCA in anxiety state group was higher than that in depression state group, While the situation in SSRIs is the opposite. More guidelines should be formulated to improve the recognition of comorbidities between specialized diseases and psychiatric disorders. Also, multi-level training should be implemented to perform the standard diagnosis and medication of mental disorders in clinical practice.

Cytotoxic quinones from the aerial parts of Morinda umbellata L.

Although Morinda umbellata L. has been used in numerous folk medicines, there is a lack of phytochemical studies on this plant. Sixteen undescribed quinones, namely, ten anthraquinones (umbellatas A-J), one naphthohydroquinone (umbellata K), one naphthohydroquinone dimer (umbellata L), and four dinaphthofuran quinones (umbellatas M-P), were isolated from the aerial parts of Morinda umbellata L. (Rubiaceae). The structures of all the isolated quinones were elucidated based on spectroscopic methods. Four of the unknown quinones (umbellatas A, H, K and M) showed potent cytotoxic effects against A431, A2780, NCI-H460, HCT116, HepG2, and MCF-7 human cancer cell lines with IC50 values of 1.3-7.1 µM. These results reveal potential lead compounds for the development of new anticancer agents.

Anti-inflammatory terpenes from Schefflera rubriflora C. J. Tseng & G. Hoo with their TNF-α and IL-6 inhibitory activities.

The 95% ethanol extract and its EtOAc and n-BuOH fractions obtained from the leaves and twigs of Schefflera rubriflora C. J. Tseng & G. Hoo showed significant inhibitory activities (33.6%, 35.7% and 40.6%, respectively) against croton oil-induced ear inflammation in mice. Bioactivity-guided isolation and separation gave eight previously undescribed terpenes or terpene glycosides. Structural elucidation was based on UV, IR, and NMR spectroscopy, MS, experimental and calculated ECD data, and Mosher's method. To identify anti-inflammatory components from the extract, all the compounds were evaluated for tumor necrosis factor-α (TNF-α) and interleukine-6 (IL-6) inhibitory activities. Four undescribed compounds inhibited mRNA expression of TNF-α and IL-6 with IC50 values of 15.3-52.4 µM.

Eight new glycosides with hepatoprotective activity isolated from the aerial parts of Morinda parvifolia.

Eight new glycosides, morindaparvins P-W, were isolated from the butanol extract of the aerial parts of Morinda parvifolia. These new structures were elucidated by comprehensive spectroscopic analysis and by comparing the experimental and calculated electronic circular dichroism spectra. The butanol extract was observed to significantly reduce the levels of alanine aminotransferase, aspartate transaminase, and lactate dehydrogenase in the sera of mice with concanavalin A-induced hepatitis. At a concentration of 10 µM, five compounds (1, and 4-7) isolated from the butanol extract possessed hepatoprotective activities against the damage induced by acetaminophen in human HepG2 liver cancer cells.

[Reviews on natural naphthoquinones and their bioactivities].

The naphthaquinones are widely distributed in plants. They are usually in higher plants, but a few of them were also found in microorganisms. There is a lot of research showing that they had multiple pharmaco-activities such as cytotoxic, antioxidant, anti-inflammatory and antibacterial activities, etc. In recent years, they have attracted extensive attention at home and abroad especially in terms of the anti-tumor activity. For further research, 69 new natural naphthoquinones reported in the last five years (2013-2017) were reviewed. They were divided into five major types: simple 1,4-naphthoquinones, furan and pyran naphthoquinones, 1,2-naphthoquinones, naphthohydroquinones and naphthoquinone polymers, which showed cytotoxic, antioxidative, anti-inflammatory and antibacterial biological activities, et al. The research of these compounds in the future was also proposed.

Glycosides of naphthohydroquinones and anthraquinones isolated from the aerial parts of Morinda parvifolia Bartl. ex DC (Rubiaceae) increase p53 mRNA expression in A2780 cells.

Eight previously undescribed naphthohydroquinone glycosides, namely morindaparvins H-O, together with four known anthraquinone glycosides were isolated from the n-BuOH extract of the aerial parts of Morinda parvifolia Bartl. ex DC (Rubiaceae). The structures of morindaparvins H-O were elucidated on the basis of spectroscopic analysis. To our knowledge, this is the first isolation of quinone glycosides from the plant M. parvifolia. The results showed that all 12 compounds at the concentration of 50 µM significantly increased p53 mRNA expression in A2780 cells compared with the blank control group.

[Constituents isolated from n-butanol extract of leaves of Moringa oleifera].

Seventeen compounds were isolated from n-butanol extract of the leaves of Moringa oleifera, using column chromatography over macroporous resin HP-20,Sephadex LH-20, and ODS. Their structures were identified as two carboline,tangutorid E(1) and tangutorid F(2); three phenolic glycosides,niazirin(3)，benzaldehyde 4-O-α-L-rhamnopyranoside(4) and 4-O-ß-D-glucopyranosidebenzoic acid(5); four chlorogenic acid and derivatives,4-caffeoylquinic acid(6),methyl 4-caffeoylquinate(7),caffeoylquinic acid(8) and methyl caffeoylquinate(9); two nucleosids,uridine(10) and adenosine(11); one flavone,quercetin 3-O-ß-D-glucopyranoside(12); five other types of compounds,phthalimidineacetic acid(13),3-pyridinecarboxamide(14),3,4-dihydroxy-benzoic acid(15),5-hydroxymethyl-2-furancarboxylic acid(16) and 5-hydroxymethyl-2-furaldehyde(17) by the spectral data of ¹H, ¹³C-NMR and MS. Among them,compounds 1-2,7,9-10,16 and 17 were isolated from M. oleifera for the first time.

A New Nortriterpenoid, a Sesquiterpene and Hepatoprotective Lignans Isolated from the Fruit of Schisandra chinensis.

A new nortriterpenoid, 19(R)-hydroxyl-wuweizidilactone H (1), and a sesquiterpene, (6R)-ß-chamigrenic acid (2), together with one known nortriterpenoid, wuweizidilactone H (3), and three known hepatoprotective lignans, micrantherin A (4), gomisin M2 (5) and schizandrin (6) were isolated from the fruit of Schisandra chinensis. Their structures were elucidated by UV, IR, HRESIMS, NMR spectra and X-ray analysis. Among them, the absolute configuration of 2 was confirmed for the first time. In vitro assays, compounds 4-6 (10 µM) exhibited hepatoprotective activities (survival rate: 44%, 43% and 44%) against damage induced by N-acetyl-p-aminophenol (APAP) in human liver carcinoma (HepG2) cells.

Phenolic Acid Profiling, Antioxidant, and Anti-Inflammatory Activities, and miRNA Regulation in the Polyphenols of 16 Blueberry Samples from China.

To investigate the anti-atherosclerosis related mechanism of blueberries, the phenolic acids (PAs) content, antioxidant and anti-inflammatory activities, as well as the microRNA (miRNA) regulation of polyphenol fractions in blueberry samples from China were studied. Sixteen batches of blueberries including 14 commercialized cultivars (Reka, Patriot, Brigitta, Bluecrop, Berkeley, Duke, Darrow, Northland, Northblue, Northcountry, Bluesource, Southgood, O'Neal, and Misty) were used in this study. Seven PAs in the polyphenol fractions from 16 blueberry samples in China were quantified by high performance liquid chromatography/tandem mass spectrometry (HPLC/MS²). The antioxidant activities of blueberry polyphenols were tested by (1,1-diphenyl-2-picrylhydrazyl [DPPH]) assay. The anti-inflammatory (tumor necrosis factor-α [TNF-α] and interleukin-6 [IL-6]) activities of the polyphenol fractions of the blueberries were investigated by using lipopolysaccharide (LPS) induced RAW 264.7 macrophages. The correlation analysis showed that the antioxidant (1,1-diphenyl-2-picrylhydrazyl [DPPH]) and anti-inflammatory (tumor necrosis factor-α [TNF-α] and interleukin-6 [IL-6]) activities of the polyphenol fractions of the blueberries were in accordance with their PA contents. Although the polyphenol-enriched fractions of blueberries could inhibit the microRNAs (miRNAs) (miR-21, miR-146a, and miR-125b) to different extents, no significant contribution from the PAs was observed. The inhibition of these miRNAs could mostly be attributed to the other compounds present in the polyphenol-enriched fraction of the blueberries. This is the first study to evaluate the PAs content, antioxidant and anti-inflammatory activities, and miRNA regulation of Chinese blueberries.

Klotho protein lowered in senile patients with brady sinus arrhythmia.

OBJECTIVE: To explore the correlationship between brady sinus arrhythmia and the levels of serum klotho protein in aged. METHODS: 104 patients over 75 years old with brady sinus arrhythmia (experiment group) were enrolled, including 34 cases of sinus arrest, 43 cases of sinus bradycardia and 25 cases of atrioventricular block. 109 patients over 75 years old without brady sinus arrhymia were chosen as control group. All subjects were monitored by Holter. The levels of serum klotho protein were detected and compared among three groups. The correlation between the frequency of sinus arrest and the levels of serum klotho protein was analyzed simultaneously. RESULTS: The levels of serum klotho protein in experiment group were lower than that in control group (P<0.01); the sinus arrest frequency was negatively correlated with the levels of serum klotho protien. The levels of serum klotho protein in patients with sinus arrest were lower than that with sinus bradycardia and atrioventricularblock (P<0.05). But there was no significant difference between sinus bradycardia group and atrioventricular block group. CONCLUSION: The levels of serum klotho protein may reflect the function of sinoatrial node and could be used as an index to estimate the function of sinoatrial node.

Correlation between serum cystatin C level and elderly hypertensive patients combined coronary heart disease.

OBJECTIVES: To explore the correlation between serum cystatin C level and elderly hypertension with coronary heart disease patients. METHODS: 500 hypertensive patients combined coronary heart disease were selected by coronary angiography. 321 of them were elderly patients with hypertension (male 204, female 117), and 400 of them were elderly patients with coronary heart disease (male 257, female 143), The serum cystatin C level of all patients were detected by immunoturbidimetry, and analyzed the correlation between the serum cystatin C level and different degree of blood pressure and the degree of coronary artery stenosis in elderly patients. RESULTS: The serum cystatin C level was closely related with the blood pressure and the degree of the coronary artery stenosis. The higher the blood pressure level and the more serious the coronary artery stenosis, the higher the serum cystatin C level; The serum cystatin C level of hypertensive patients with coronary heart disease patients (Group D) were markedly higher than the level of the patients without hypertension and coronary heart disease patients (Group A), and the level of the patients with coronary heart disease (Group B) and the hypertension group (Group C) (P < 0.05). CONCLUSION: The serum cystatin C level of elderly patients with hypertension and coronary heart disease were closely related with the degree of blood pressure and coronary arteries stenosis. The serum cystatin C maybe a predictor of disease severity in elderly hypertensive patients with coronary heart disease.

ABCA1 mRNA expression and cholesterol outflow in U937 cells.

OBJECTIVE: To investigate the oxidized low density lipoprotein (oxLDL) on U937 cell ATP-binding cassette transporter A1 (ABCA1) mRNA expression and cholesterol efflux situation. METHODS: Human U937 cells were incubated with gradient concentrations of oxLDL (0, 25, 50, 75, 100, 125 mg/L), and then dyed by oil red O to estimate the content of intracelluar lipid and detect the expressing quantity of ABCA1 mRNA by Real-time Fluorescence quantitative PCR simultaneously. Calculating the cholesterol efflux rates by using the scintillation counter to detect the amount of H(3)-cholesterol in each well cell culture plate and medium. RESULTS: Real-time Fluorescence quantitative PCR analysis showed that the expression levels of ABCA1 mRNA in monocytes were lower than basal line when not intervened with oxLDL, and increased drastically with oxLDL stimulation, significant difference compared with controls (P<0.01), and reached the highest level at oxLDL 50 mg/L, nevertheless, continuously increasing the concentration of oxLDL above 50 mg/L, the expression decreased. So is the outflowing rate of intracelluar lipid. Oil red O dyeing results also suggested that celluar lipid content was the highest when intervened with 125 mg/L oxLDL, and increased most obviously at 50 mg/L oxLDL. Cholesterol outflow result also demonstrated that cholesterol outflow rate related with the ABCA1 mRNA expressing quantity. CONCLUSION: With the increase of intervening concentration of oxLDL on U937cells, the exprssion of ABCAl mRNA represented that rising before 50 mg/L oxLDL, and then decreasing, reaching the top point at 50 mg/L oxLDL. So was the change in the outflowing rate of intracelluar lipid.

Prediction of Metabolic Gene Biomarkers for Neurodegenerative Disease by an Integrated Network-Based Approach.

Neurodegenerative diseases (NDs), such as Parkinson's disease (PD) and Huntington's disease (HD), have become more and more common among aged people worldwide. One hallmark of NDs is the presence of intracellular accumulation of specific pathogenic proteins that may result from abnormal function of metabolic processes. Previously, we have developed a computational method named Met-express that predicted key enzyme-coding genes in cancer development by integrating cancer gene coexpression network with the metabolic network. Here, we applied Met-express to predict key enzyme-coding genes in both PD and HD. Functional enrichment analysis and literature review of predicted genes suggested that there might be some common pathogenic metabolic pathways for PD and HD. We further found that the predicted genes had significant functional association with known disease genes, with some of them already documented as biomarkers or therapeutic targets for NDs. As such, the predicted metabolic genes may be of use as novel biomarkers not only for ND diagnosis but also for potential therapeutic treatments.

Vasorelaxant activities of Danhong injection and their differential effects on the rat abdominal aorta and mesenteric artery.

Previous studies have found that Danhong injection (DHI), an extensively used herbal extract preparation in China, might be a powerful vasodilator. The aims of this study were to determine the vascular activity of DHI and its effects on arteries of different sizes. The results showed that DHI significantly inhibited rat-hindquarters and rabbit-ear vasoconstriction elicited by norepinephrine (NE) perfusion and markedly relaxed KCl-contracted and NE-contracted rat abdominal aortic and mesenteric artery rings. The endothelium made only a minor contribution to the vasorelaxant effect of DHI on artery segments. The vasorelaxant effect of DHI varied with the artery size, with larger arteries exhibiting a more sensitive and potent vasodilator response. DHI relaxed NE-induced vasoconstriction probably through inhibition of the intracellular Ca2+ release through the inositol triphosphate receptor system in the abdominal aorta and mesenteric artery, along with blockage of extracellular Ca2+ influx through the receptor-linked Ca2+ channels in the mesenteric artery. In addition, DHI completely relaxed KCl-induced contraction in both of the arteries, suggesting that inhibition of Ca2+ influx through voltage-gated Ca2+ channels is involved in the vasorelaxant effect of DHI. This elucidation of the vascular effects of DHI and the underlying mechanisms could lead to improved clinical applications.

α-Klotho is an acute phase protein and altered by restraint stress in mice.

OBJECTIVE: To identify whether α-Klotho is a kind of acute phase protein and alternation in the level of protein and mRNA under restraint stress. METHOD: 48 mice were divided into three groups of 16: 1h, 10h and 20 h treat time group. Each group included restraint stress and control subgroup with same number animals (n = 8). ELISA was utilized in the assay of serum α-Klotho and corticosterone, RT-PCR was occupied in the detection of the expression of α-Klotho mRNA in renal tissue. RESULT: α-Klotho protein concentration of control group 1h, 10h and 20 h was 757.71 ± 333.93 pg/ml, 687.38 ± 342.79 pg/ml and 912.90 ± 337.8 pg/ml, respectively. While the concentration of restraint stress group 1h, 10h and 20 h was 726.40 ± 342.79 pg/ml, 1261.54 ± 442.71 pg/ml, and 1696.18 ± 404.11 pg/ml. There was no significant difference among 1h, 10h and 20 h control groups (P > 0.05) as well as 1h treat time subgroup. Compared with respective control group, the difference of restraint stress group in 10h and 20 h treat time group was significant (P < 0.05). The expression of α-Klotho mRNA was slightly downregulated even when the mice underwent a 1 hour restraint stress, though without significance (p > 0.05). Prominent fold change, 2.02 and 2.46, happened in 10h and 20 h restraint group with significance (P < 0.05 and P < 0.01), respectively. CONCLUSION: α-Klotho is a kind of acute phase protein. The serum α-Klotho protein is promoted while the α-Klotho mRNA is downregulated under the constraint stress.