Intestinal epithelial Krüppel-like factor 4 alleviates endotoxemia and atherosclerosis through improving NF-κB/miR-34a-mediated intestinal permeability.

Maintenance of intestinal barrier function contributes to gastrointestinal homeostasis and therefore cardiovascular diseases. A number of studies show that intestinal permeability is affected by excessive inflammatory responses. Krüppel-like factor (KLF) 4 is one of the critical transcriptional factors, which controls multiple immune responses. In this study we investigated the role of KLF4 in regulating intestinal inflammation and permeability during the atherosclerotic process. Atherosclerotic model was established in ApoE-/- mice by feeding a high fat high cholesterol (HFHC) diet. We showed that colon expression levels of KLF4 and tight junction proteins were significantly decreased whereas inflammatory responses increased in atherosclerotic mice. Overexpression of colon epithelial Klf4 decreased atherosclerotic plaque formation and vascular inflammation in atherosclerotic mice, accompanied by remarkable suppression of intestinal NF-κB activation. We found that overexpression of epithelial Klf4 in atherosclerotic mice significantly increased intestinal tight junction expression and ameliorated endotoxemia, whereas replenishment of LPS abolished these benefits. Overexpression of Klf4 reversed LPS-induced permeability and downregulation of ZO-1 and Occludin in Caco-2 cells in vitro. HFHC diet stimulated the expression of epithelial microRNA-34a, whereas silence of epithelial Klf4 abolished the benefits of microRNA-34a sponge, a specific miR-34a inhibitor, on intestinal permeability and atherosclerotic development. A clinical cohort of 24 atherosclerotic patients supported colon KLF4/NF-κB/tight junction protein axis mediated intestine/cardiovascular interaction in patients with atherosclerosis. Taken together, intestinal epithelial KLF4 protects against intestinal inflammation and barrier dysfunction, ameliorating atherosclerotic plaque formation.

Bmal1 inhibits phenotypic transformation of hepatic stellate cells in liver fibrosis via IDH1/α-KG-mediated glycolysis.

Hepatic stellate cells (HSCs) play an important role in the initiation and development of liver fibrogenesis, and abnormal glucose metabolism is increasingly being considered a crucial factor controlling phenotypic transformation in HSCs. However, the role of the factors affecting glycolysis in HSCs in the experimental models of liver fibrosis has not been completely elucidated. In this study, we showed that glycolysis was significantly enhanced, while the expression of brain and muscle arnt-like protein-1 (Bmal1) was downregulated in fibrotic liver tissues of mice, primary HSCs, and transforming growth factor-ß1 (TGF-ß1)-induced LX2 cells. Overexpression of Bmal1 in TGF-ß1-induced LX2 cells blocked glycolysis and inhibited the proliferation and phenotypic transformation of activated HSCs. We further confirmed the protective effect of Bmal1 in liver fibrosis by overexpressing Bmal1 from hepatic adeno-associated virus 8 in mice. In addition, we also showed that the regulation of glycolysis by Bmal1 is mediated by the isocitrate dehydrogenase 1/α-ketoglutarate (IDH1/α-KG) pathway. Collectively, our results indicated that a novel Bmal1-IDH1/α-KG axis may be involved in regulating glycolysis of activated HSCs and might hence be used as a therapeutic target for alleviating liver fibrosis.

The Relationship between Erythrocyte Sedimentation Rate and Erythrocyte Sedimentation/Red Blood Cell Ratio and Disease Activity in Systemic Lupus Erythematosus.

BACKGROUND: The treatment options of systemic lupus erythematosus (SLE) patients in active and inactive phases are very different clinically, and the prognosis of patients with active SLE is much worse than inactive patients. However, the present indicators for diagnosis of SLE in activity are limited and inefficient. METHODS: Three hundred thirty patients with SLE were included. All patients are classified as SLEDAI (systemic lupus erythematosus disease activity index) > 4 as active and SLEDAI ≤ 4 as inactive. The linear correlation between variables was assessed by Pearson's correlation analysis. The difference between parameters in active and inactive patients was evaluated by the Mann-Whitney U test. The evaluation capacity of erythrocyte sedimenta-tion/red blood cell (ERR) and red blood cell/albumin ratio (RAR) on SLE activity was determined by bivariate regression analysis. Sensitivity and specificity are assessed by receiver operating characteristic curve (ROC). RESULTS: Compared with the inactive SLE, ESR (52.97 ± 35.66 vs. 32.38 ± 29.16 p < 0.001), ERR (15.40 ± 12.41 vs. 8.19 ± 8.10 p < 0.001) and RAR (0.13 ± 0.10 vs. 0.11 ± 0.20 p = 0.038) are all elevated in active SLE (52.97 ± 35.66 vs. 32.38 ±2 9.16 p < 0.001). ERR shows better correlation than RAR with ESR (p < 0.001 vs. p = 0.911). Patients with active SLE exhibited higher SLEDAI than those with inactive SLE (8.67 ± 2.67 vs. 3.27 ± 1.36, p < 0.001). According to ROC analysis, when ESR levels > 58.5 and ERR levels > 13.18, the sensitivity is 37.6% and 45.2%, the specificity is 83.0% and 83.2%. CONCLUSIONS: ESR and ERR are potential indicators for diagnosis of active and inactive SLE.

Orthodontic-surgical treatment for severe skeletal class II malocclusion with vertical maxillary excess and four premolars extraction: A case report.

BACKGROUND: Patient satisfaction with facial appearance at the end of orthodontic camouflage treatment is very important, especially for skeletal malocclusion. This case report highlights the importance of the treatment plan for a patient initially treated with four-premolar-extraction camouflage, despite indications for orthognathic surgery. CASE SUMMARY: A 23-year-old male sought treatment complaining about his unsatisfactory facial appearance. His maxillary first premolars and mandibular second premolars had been extracted, and a fixed appliance had been used to retract his anterior teeth for two years without improvement. He had a convex profile, a gummy smile, lip incompetence, inadequate maxillary incisor inclination, and almost a class I molar relationship. Cephalometric analysis showed severe skeletal class II malocclusion (A point-nasion-B point = 11.5°) with a retrognathic mandible (sella-nasion-B point = 75.9°), a protruded maxilla (sella-nasion-A point = 87.4°), and vertical maxillary excess (upper incisor to palatal plane = 33.2 mm). The excessive lingual inclination of the maxillary incisors (upper incisor to nasion-A point line = -5.5°) was due to previous treatment attempts to compensate for the skeletal class II malocclusion. The patient was successfully retreated with decompensating orthodontic treatment combined with orthognathic surgery. The maxillary incisors were repositioned and proclined in the alveolar bone, the overjet was increased, and a space was created for orthognathic surgery, including maxillary impaction, anterior maxillary back-setting, and bilateral sagittal split ramus osteotomy to correct his skeletal anteroposterior discrepancy. Gingival display was reduced, and lip competence was restored. In addition, the results remained stable after 2 years. The patient was satisfied with his new profile as well as with the functional malocclusion at the end of treatment. CONCLUSION: This case report provides orthodontists a good example of how to treat an adult with severe skeletal class II malocclusion with vertical maxillary excess after an unsatisfactory orthodontic camouflage treatment. Orthodontic and orthognathic treatment can significantly correct a patient's facial appearance.

Coordination effect enhanced visualization of latent fingerprint with Eu (TTA)3phen-SiO2 microspheres.

Latent fingerprint (LFP) powders are crucial in the detection of LFPs in forensic science. However, it is often plagued by poor image resolution and low contrast. Herein, enhanced LFP fluorescence (FL) visualizations are achieved by doping Eu(III) coordination compound Eu(TTA)3phen directly into SiO2 microspheres instead of Eu(III) ions. Using the synthesized Eu(TTA)3phen-SiO2 microspheres, the fine characteristic structure of LFP can be seen and recognized under 365 nm irradiation, up to Level 3. However, the Eu3+-SiO2 microspheres were difficult to recognize the Level 2,3 fingerprint structure. The difference between the ridge and furrow gray values of Eu(TTA)3phen-SiO2 microspheres is 2.1 times that of Eu3+-SiO2 microspheres. The coordination effect increased the asymmetry around Eu(III) ions, resulting in the ultrasensitive 5D0→7F2 transition, thus increasing the FL intensity, and the uniform doping of the Eu(III) coordination compound into SiO2 also reduced the surface FL quenching due to shielding from oxygen. Under this dual effect, the LFP performance of Eu(TTA)3phen-SiO2 microspheres has been significantly improved. We believe that this novel and easy LFP visualization method is a promising routine in specific target detection including criminal investigation, customhouse check-in, and drug control.

Epididymis cell atlas in a patient with a sex development disorder and a novel NR5A1 gene mutation.

This study aims to characterize the cell atlas of the epididymis derived from a 46,XY disorders of sex development (DSD) patient with a novel heterozygous mutation of the nuclear receptor subfamily 5 group A member 1 (NR5A1) gene. Next-generation sequencing found a heterozygous c.124C>G mutation in NR5A1 that resulted in a p.Q42E missense mutation in the conserved DNA-binding domain of NR5A1. The patient demonstrated feminization of external genitalia and Tanner stage 1 breast development. The surgical procedure revealed a morphologically normal epididymis and vas deferens but a dysplastic testis. Microfluidic-based single-cell RNA sequencing (scRNA-seq) analysis found that the fibroblast cells were significantly increased (approximately 46.5%), whereas the number of main epididymal epithelial cells (approximately 9.2%), such as principal cells and basal cells, was dramatically decreased. Bioinformatics analysis of cell-cell communications and gene regulatory networks at the single-cell level inferred that epididymal epithelial cell loss and fibroblast occupation are associated with the epithelial-to-mesenchymal transition (EMT) process. The present study provides a cell atlas of the epididymis of a patient with 46,XY DSD and serves as an important resource for understanding the pathophysiology of DSD.

Hesitate between confocal laser endomicroscopy and narrow-band imaging: how to choose a better method in the detection of focal precancerous state of gastric cancer.

BACKGROUND: With a high incidence globally, deaths form gastric cancer (GC) are not rare. Early diagnosis is crucial to ameliorate its prognosis. Confocal laser endomicroscopy (CLE) and narrow band imaging (NBI) have been extensively applied in gastroscopy, particularly when it comes to the detection and management of premalignant gastric lesion. Our meta-analysis intends to appraise the diagnostic capability and compare the efficacy of NBI and CLE for focal precancerous state of gastric cancer. METHODS: We performed a literature search up to November 5, 2020 in online databases and major conferences. Two investigators assessed the methodological bias by QUADAS-2, followed by sophisticated study selection and data exaction to make a comparison between sensitivity, specificity, positive and negative likelihood values, and diagnostic odds ratio. A symmetric summary receiver-operating curve (sROC) and its area under the curve (AUC) were used to estimate threshold effect. Additionally, we evaluated the publication bias by Deeks' asymmetry test. RESULTS AND CONCLUSIONS: Four studies involved 248 patients and 526 lesions. In analysis drawn from every lesion, the NBI's pooled sensitivity and specificity were 87% (95% CI: 0.80-0.92) and 85% (95% CI: 0.75-0.91), and those of CLE were 90% (95% CI: 0.85-0.91) and 87% (95% CI: 0.83-0.91). CLE illustrated that the pooled two were slightly higher than NBI when compared at the level of every lesion. The AUC for NBI and CLE was 0.92 (0.90-0.94) and 0.95 (0.92-0.96), and there might be a threshold effect, according to the shoulder-like distribution of scatter points in the sROC. We did not find obvious publication bias in our meta-analysis.

[Predictive value of human fatty acid binding protein for myocardial ischemia and injury in perioperative period of cardiac surgery].

OBJECTIVE: To evaluate the value of human fatty acid binding protein (h-FABP) in predicting myocardial ischemia and injury in the perioperative period of cardiac surgery, we observed the dynamic changes of h-FABP in perioperative period of patients underwent coronary artery bypass grafting and ventricular septal defects repairing surgery, and evaluated the relationship of h-FABP and ischemia modified albumin (IMA), CK-MB, cTnI. METHODS: Patients underwent coronary artery bypass grafting (n=30) and ventricular septal defect repairing (n=30) surgery between February 2008 and December 2008 were included in this study. Venous blood sample was obtained at preoperative, aortic clamping, aortic unclamping of 10 min, 2 h, 6 h, 12 h, 24 h for the measurements of h-FABP, IMA, cTnI and CK-MB. RESULTS: h-FABP and IMA changed in the same way at various examined time points, h-FABP changes also paralleled cTnI and CK-MB changes, h-FABP peaked early during myocardial ischemia and injury and returned to baseline level at 2 h post myocardial ischemia and injury. Linear correlation analysis showed that the peak value of h-FABP was positively correlated with IMA, CK-MB and cTnI in both CABG group (r = 0.948, 0.964 and 0.961, P < 0.05) and in the VSD group (r = 0.986, 0.978 and 0.957). CONCLUSIONS: h-FABP is an early diagnostic parameter reflecting perioperative myocardial ischemia and injury in cardiac surgery. Quantitative h-FABP monitoring could predict the severity of myocardial ischemia and injury early during cardiac surgery.

Melatonin Reduces Androgen Production and Upregulates Heme Oxygenase-1 Expression in Granulosa Cells from PCOS Patients with Hypoestrogenia and Hyperandrogenia.

BACKGROUND/AIMS: Polycystic ovary syndrome (PCOS) is an endocrine disorder characterized by abnormal hormone levels in peripheral blood and poor-quality oocytes. PCOS is a pathophysiological syndrome caused by chronic inflammation and oxidative stress. The aim of this study was to investigate the mechanism of melatonin regulation on androgen production and antioxidative damage in granulosa cells from PCOS patients with hypoestrogenia and hyperandrogenia. METHODS: Cumulus-oocyte complexes were collected from PCOS patients who had low levels of estrogen in follicular fluids. RESULTS: Melatonin triggered upregulation of cytochrome P450 family 19 subfamily A member 1 (CYP19A1) expression via the extracellular signal-regulated kinase pathway in luteinized granulosa cells. As a result, conversion of androgen to 17ß-estradiol was accelerated. We also found that melatonin significantly reduced the levels of inducible nitric oxide (NO) synthetase and NO in luteinized granulosa cells. Levels of transcripts encoding NF-E2-related factor-2 and its downstream target heme oxygenase-1 were also increased, leading to anti-inflammatory and antioxidant effects. We also found that melatonin could improve oocyte development potential. CONCLUSION: Our preliminary results showed that melatonin had a positive impact on oocyte quality in PCOS patients with hypoestrogenia and hyperandrogenia.

[Effect of Nutrient Loadings on the Regulation of Water Nitrogen and Phosphorus by Vallisneria natans and Its Photosynthetic Fluorescence Characteristics].

Submerged macrophytes are an important component of aquatic ecosystems. During the growing period, submerged macrophytes can absorb nitrogen and phosphorus nutrients to reduce pollution loadings. Shoots of submerged macrophytes can also promote the adhesion of suspended substances in water, reducing the turbidity. The release of nutrients in sediments can be suppressed by its root system, and the resuspension of sediments caused by disturbance of winds and waves can also be resisted. The role of submerged macrophytes in ecological restoration of eutrophic lakes has attracted widespread attention. In 1960, the submerged plants Vallisneria natans and Potamogeton malaianus had been the dominant species in East Taihu. However after 2002, Nymphoides peltatum, Elodea nattalii, P. malaianus, etc. have gradually taken over the dominant roles along with significant elevations of nitrogen and phosphorus levels. Nutrients in water are not the only key factors causing eutrophication of water bodies; the nutrient source for submerged plant growth affect both the purification efficiency and the photosynthetic characteristics of submerged macrophytes. Excessive nitrogen and phosphorus concentrations can inhibit the photosynthetic physiological activities of submerged macrophytes, affecting the succession of aquatic vegetation. In addition, under high nutrient conditions, the competition from periphytic algae and planktonic algae may also directly poison submerged macrophytes, leading to its degradation and disappearance. Systematic studies on the regulation and photosynthetic fluorescence response mechanism of submerged macrophytes to varied nutrient loadings are helpful in revealing their relationships. The seedlings of submerged macrophyte V. natans were transplanted in a laboratory mesocosm to study the effect of nutrient loadings on its regulation of water nitrogen and phosphorus. Three nitrogen and phosphorus loadings from low, medium, and high levels derived from nitrate, ammonium, and phosphate were setup as the aquatic medium for the plant growth. Twelve harvests were carried out to determine the evolution of nutrient removal performance of V. natans. Its photosynthetic fluorescence characteristics were measured by a pulse-amplitude modulated fluorometer (Diving-PAM). Results showed that the nitrogen and phosphorus adsorption abilities of V. natans were gradually enhanced with the increase of nutrient concentrations in the range of TN ≤ 12 mg·L-1 and TP ≤ 1.0 mg·L-1. In the treatment of high nutrient concentrations (TN=12 mg·L-1 and TP=1.0 mg·L-1), the removal rates of nitrogen and phosphorus reached more than 95%. V. natans preferentially absorbed ammonium nitrogen when its concentration was high. The medium nutrient concentrations (TN:8-12 mg·L-1 and TP:0.6-1.0 mg·L-1) did not significantly affect the Fv/Fm ratio of leaves. However, the low nutrient concentrations (TN=3 mg·L-1 and TP=0.3 mg·L-1) could improve the Fv/Fm ratio of leaves and were beneficial for the growth of V. natans. The inhibition of photosynthetic activity and light tolerance were enhanced with the increase in nutrient concentrations. The photosynthetic activity of V. natans gradually recovered with no significant changes in the capacity for light harvesting, when the nutrient concentrations gradually decreased in the water. Our results indicate that the high nitrogen and phosphorus loadings indeed hamper the photosynthetic capacity, which may subsequently restrain the maintenance of the dominance of V. natans in the submerged macrophyte communities.

Human erythrocyte lifespan measured by Levitt's CO breath test with newly developed automatic instrument.

Existing standard techniques for erythrocyte (RBC) lifespan measurement, such as quantitation of labeling with isotopes or biotin, are cumbersome and time-consuming. Given that endogenous CO originates mainly from degraded RBCs, a team lead by Levitt developed a CO breath test to enable more efficient RBC lifespan estimation. The purpose of this study was to evaluate the reliability of Levitt's CO breath test method with our newly developed automatic instrument. RBC lifespan measurements conducted by Levitt's CO breath test method were conducted in 109 healthy subjects and 91 patients with chronic hemolytic anemia. In healthy subjects, the RBC lifespan was 126 ± 26 days, similar to values obtained with classical standard labeling methods. RBC lifespan did not differ significantly between males and females or between juveniles and adults, and did not correlate with age. To our knowledge, this datum represents an RBC lifespan average for the largest sample to date. In subjects with hemolytic anemia, RBC lifespan was 29 ± 14 days, which is significantly shorter than that of the healthy subjects (p = 0.001). Using 75 days as a cut-off, diagnostic accuracy for hemolytic anemia in the present study sample was 100%. In conclusion, the present results indicate that Levitt's CO breath test is an ideal method for human RBC lifespan measurement, and the newly developed automatic instrument is reliable and convenient for clinical practice.

Development of a novel ultrasound-assisted surfactant-enhanced emulsification microextraction method and its application to the analysis of eleven polycyclic aromatic hydrocarbons at trace levels in water.

A novel ultrasound-assisted surfactant-enhanced emulsification microextraction (UASEME) technique has been proposed by using low-density extraction solvents. In the proposed technique, Tween 80 and cyclohexane were injected into 5-mL glass test tubes with conical bottoms, containing 5.00 mL of a water sample that was located inside the ultrasonic bath. When the extraction process was finished, the glass test tube was sealed with a rubber plug and then placed upside down in a centrifuge. The finely dispersed droplets of cyclohexane collected at the conical bottom of test tube because the density of cyclohexane is less than of water, and the PAHs were concentrated in the cyclohexane. Next, 5 µL of the cyclohexane that collected at the conical bottom was removed using a 10-µL microsyringe and injected into high performance liquid chromatography coupled with fluorescence detection (HPLC-FLD) for analysis. The proposed method avoided the use of chlorinated solvents, which have been widely used as extraction solvents in a normal UASEME assay. Parameters that affected the extraction efficiency, such as the type and volume of the extraction solvent, the type and concentration of the surfactant, and the ultrasound emulsification time and salt addition, were investigated and optimised for the method. Under the optimum conditions, the enrichment factors ranged between 90 and 247. The limits of detection of the method were 0.6-62.5 ng L(-1). Good recoveries and repeatability of the method for the eleven PAHs were also obtained. The proposed UASEME technique has been demonstrated to be simple, practical and environmentally friendly for the determination of PAH residues in real water samples.

Quantification of multi-residue levels in peach juices, pulps and peels using dispersive liquid-liquid microextraction based on floating organic droplet coupled with gas chromatography-electron capture detection.

In this paper, polychlorinated biphenyl (PCB), organochlorine pesticide (OCP) and pyrethroid pesticides in peach was investigated by comparing their residual level in peach juice, pulps and peels using dispersive liquid-liquid microextraction based on solidification of floating organic droplet (DLLME-SFO) combined with gas chromatography-electron capture detection (GC-ECD). Extraction conditions such as the type of extractant, volume of extractant and dispersant, salt effect and extraction time were optimized. For juice samples, the linearity of the method was obtained in the range of 10-2000 ng L(-1),with determination coefficients>0.99. The limits of detection (LOD) of the method were ranged between 2.8 and 18.5 ng L(-1). For pulp and peel samples, the developed method is linear over the range assayed, 1-20 µg kg(-1),with coefficients also >0.99. The relative recoveries of compounds analyzed from juice, pulp and peel samples were in the range of 73-106% with a relative standard deviation between 2.6 and 11.8%. The proposed method was applied to the simultaneous analysis of residues in real peach juice, pulp and peel samples. As a result, there were no target analytes found in peach juices and pulps while 3.3 µg kg(-1) cyhalothrin and 3.5 µg kg(-1) fenvalerate were found in peels. The experiment results revealed that the pyrethroid residues just deposited on the peels of the fruits, but did not move into pulps and juices.