Nebivolol, an antihypertensive agent, has new application in inhibiting melanoma.

Repurposing existing drugs for cancer therapy has become an important strategy because of its advantages, such as cost reduction, effect and safety. The present study was designed to investigate the antimelanoma effect and possible mechanisms of action of nebivolol, which is an approved and widely prescribed antihypertensive agent. In this study, we explored the effect of nebivolol on cell proliferation and cell activity in melanoma in vitro and the potential antimelanoma mechanism of nebivolol through a series of experiments, including the analysis of the effects with regard to cell apoptosis and metastasis. Furthermore, we evaluated the antimelanoma effect on xenograft tumor models and inspected the antimelanoma mechanism of nebivolol in vivo using immunohistochemical and immunofluorescence staining assays. As results in this work, in vitro , nebivolol possessed a strong activity for suppression proliferation and cell cycle arrest on melanoma. Moreover, nebivolol significantly induced cell apoptosis in melanoma through a mitochondrial-mediated endogenous apoptosis pathway. Additionally, nebivolol inhibited melanoma cell metastasis. More importantly, nebivolol exhibited significantly effective melanoma xenograft models in vivo , which related to the mechanism of apoptosis induction, proliferation inhibition, metastasis blocking and angiogenesis arrest. Overall, the data of the present study recommend that nebivolol holds great potential in application as a novel agent for the treatment of melanoma.

Epigallocatechin-3-gallate ameliorates lipopolysaccharide-induced acute thymus involution in mice via AMPK/Sirt1 pathway.

The thymus, a site to culture the naïve T lymphocytes, is susceptible to atrophy or involution due to aging, inflammation, and oxidation. Epigallocatechin-3-gallate (EGCG) has been proven to possess anti-inflammatory, antioxidant, and antitumor activity. Here, we investigate the effects of EGCG on thymic involution induced by lipopolysaccharide (LPS), an endotoxin derived from Gram-negative bacteria. The methodology included an in vivo experiment on female Kunming mice exposed to LPS and EGCG. Morphological assessment of thymic involution, immunohistochemical detection, and thymocyte subsets analysis by flow cytometry were further carried out to evaluate the potential role of EGCG on the thymus. As a result, we found that EGCG alleviated LPS-induced thymic atrophy, increased mitochondrial membrane potential and superoxide dismutase levels, and decreased malondialdehyde and reactive oxygen species levels. In addition, EGCG pre-supplement restored the ratio of thymocyte subsets, the expression of autoimmune regulator, sex-determining region Y-box 2, and Nanog homebox, and reduced the number of senescent cells and collagen fiber deposition. Western blotting results indicated that EGCG treatment elevated LPS-induced decrease in pAMPK, Sirt1 protein expression. Collectively, EGCG relieved thymus architecture and function damaged by LPS via regulation of AMPK/Sirt1 signaling pathway. Our findings may provide a new strategy on protection of thymus from involution caused by LPS by using EGCG. And EGCG might be considered as a potential agent for the prevention and treatment of thymic involution.

Potential mechanism of pyrotinib-induced diarrhea was explored by gut microbiome and ileum metabolomics.

Pyrotinib is a novel epidermal growth factor receptor/human epidermal growth factor receptor-2 (HER2) tyrosine kinase inhibitor that exhibited clinical efficacy in patients with HER2-positive breast cancer and HER2-mutant/amplified lung cancer. However, severe diarrhea adverse responses preclude its practical use. At present, the mechanism of pyrotinib-induced diarrhea is unknown and needs further study. First, to develop a suitable and reproducible animal model, we compared the effects of different doses of pyrotinib (20, 40, 60 and 80 mg/kg) in Wistar rats. Second, we used this model to examine the intestinal toxicity of pyrotinib. Finally, the mechanism underlying pyrotinib-induced diarrhea was fully studied using gut microbiome and host intestinal tissue metabolomics profiling. Reproducible diarrhea occurred in rats when they were given an 80 mg/kg daily dose of pyrotinib. Using the pyrotinib-induced model, we observed that Lachnospiraceae and Acidaminococcaceae decreased in the pyrotinib groups, whereas Enterobacteriaceae, Helicobacteraceae and Clostridiaceae increased at the family level by 16S rRNA gene sequence. Multiple bioinformatics methods revealed that glycocholic acid, ursodeoxycholic acid and cyclic AMP increased in the pyrotinib groups, whereas kynurenic acid decreased, which may be related to the pathogenesis of pyrotinib-induced diarrhea. Additionally, pyrotinib-induced diarrhea may be associated with a number of metabolic changes mediated by the gut microbiome, such as Primary bile acid biosynthesis. We reported the establishment of a reproducible pyrotinib-induced animal model for the first time. Furthermore, we concluded from this experiment that gut microbiome imbalance and changes in related metabolites are significant contributors to pyrotinib-induced diarrhea.

Synthesis of novel oxazol-5-one derivatives containing chiral trifluoromethyl and isoxazole moieties as potent antitumor agents and the mechanism investigation.

In this study, a series of novel oxazol-5-one derivatives containing a chiral trifluoromethyl and isoxazole moiety were synthesized and evaluated for cytotoxic activities. Among them, 5t was the most effective compound against HepG2 liver cancer cells with an IC50 of 1.8 µM. 5t inhibited cell proliferation, migration, invasion, and induced cell cycle arrest and apoptosis in vitro. Nevertheless, the potential anti-hepatocellular carcinoma (HCC) target and mechanism of 5t were unclear. This work aimed to seek the molecular target of 5t against HCC and investigate its mechanism. Liquid chromatography tandem-mass spectrometry was used to identify peroxiredoxin 1(PRDX1) as a possible target of 5t. Cellular thermal shift assay, drug affinity responsive target stability, and molecular docking provided conclusive evidence that 5t targeted PRDX1 and inhibited its enzymatic activity. 5t augmented the level of reactive oxygen species (ROS) and led to ROS-dependent DNA damage, endoplasmic reticulum stress, mitochondrial dysfunction, and apoptosis in HepG2 cells. Silencing PRDX1 also resulted in ROS-mediated apoptosis in HepG2 cells. In vivo, 5t inhibited mouse tumor growth by increasing oxidative stress. Briefly, our studies revealed that compound 5t targeted PRDX1 through a ROS-dependent mechanism, highlighting the future development of compound 5t as a novel therapeutic drug for HCC.

Association between sun-protective behaviors and hypertension: a cross-sectional study from National Health and Nutrition Examination Survey 2009 to 2014.

BACKGROUND: In previous studies, sun-protective behaviors increased cardiovascular incidence. Our present article is to further analyze the potential relationship between sun-protective behaviors (staying in the shade, wearing long-sleeved clothing, and applying sunscreen) and hypertension. METHOD: The present cross-sectional study evaluated 8,613 participants (aged 20-60 years) from the National Health and Nutrition Examination Survey (NHANES) obtained between 2009 and 2014. We performed multiple logistic regression analysis to examine the relationship between sun-protective behaviors and hypertension. Subgroup analysis was then performed. Multiple linear regression analysis was utilized to examine the relationship of sun-protective behaviors and each sun-protective behavior with systolic and diastolic blood pressure, stratified by sex and race. RESULTS: A total of 8,613 participants (weighted n = 127,909,475) were applied in our study, including 1,694 hypertensive subjects. Our study demonstrated that sun-protective behaviors of the 2-3 category were associated with increased risk of hypertension, but not with higher systolic and diastolic blood pressure. In subgroup analysis, men, Mexican American, and 25 < BMI ≤ 30 who reported sun-protective behaviors (2-3) were prone to hypertension. Multiple linear regression models showed that non-Hispanic white men with sun-protective behaviors (2-3) were positively associated with systolic and diastolic blood pressure. The association between other-Hispanic men with frequent wearing long-sleeved clothing and diastolic blood pressure was positively correlated. CONCLUSION: Sun-protective behaviors of the 2-3 category could increase the incidence of hypertension, but not increase systolic and diastolic blood pressure. We only found that non-Hispanic white men who reported sun-protective behaviors (2-3) were positively associated with systolic and diastolic blood pressure. These findings suggested that excessive sun-protective behaviors should be avoided.

Nomogram-based development and evaluation for predictions of 30-day and 1-year survival in patients with spontaneously ruptured hepatocellular carcinoma.

BACKGROUND: Accurately predicting the prognosis of patients with spontaneously ruptured hepatocellular carcinoma (HCC) is crucial for effective clinical management. The aim of the present study was to establish and evaluate prediction models for 30-day and 1-year survival in patients with spontaneously ruptured HCC. METHODS: A total of 118 patients with spontaneous rupture HCC were enrolled. Univariate and multivariate analyses were performed using logistic-regression model and Cox proportional-hazard model. The identified indicators were used to establish prediction models, the performance of which we compared with those of commonly used liver disease scoring models. The survival possibilities of different risk categories were calculated using the newly developed models. RESULTS: Largest tumor size (LTS), serum albumin (ALB), total bilirubin (TBil), and serum creatinine were identified as independent predictors, which were used to establish a 30-day survival prediction model. LTS, BCLC staging, ALB, TBil, hepatectomy at rupture, and TACE during follow-up were identified as independent predictors of 1-year survival model. The 30-day survival model had sensitivity of 79.3%, specificity of 87.1%, and an AUC of 0.879, exhibiting better predictive performance than scores for Chronic Liver Failure Consortium Acute Decompensation score (CLIF-C ADs) and Model for End-stage Liver Disease (MELD). The 1-year survival model had sensitivity of 66.7%, specificity of 94.6%, and an AUC of 0.835, showing better predictive performance than Albumin-Bilirubin (ALBI), Child-Pugh, CLIF-C ADs, and MELD. After stratification, survival possibilities were 90.9 and 21.1% in low- and high-risk groups within 30 days, respectively, and 43.90, 4.35%, and 0 in low-, intermediate-, and high-risk groups at 1 year, respectively. CONCLUSIONS: The established models exhibited good performance in predicting both 30-day and 1-year survival in patients with spontaneously ruptured HCC.

Radioiodinated hypericin as a tracer for detection of acute myocardial infarction: SPECT-CT imaging in a swine model.

PURPOSE: Hypericin (Hyp) is a natural compound with a newly discovered necrosis-avidity, which can be exploited as a necrosis-avid tracer once labeled with radioactive iodine as has been tested in rodent models. This study was to evaluate the effect of radioiodinated Hyp (131I-Hyp) for imaging detection of acute myocardial infarction (AMI) in conditions closer to clinical scenarios. METHODS: We established swine AMI models (n = 6) which were intravenously given 131I-Hyp and 99mTc-sestamibi and underwent SPECT-CT imaging with high- and low-energy collimators. The acquired SPECT images were fused with cardiac CT images and correlated with postmortem autoradiography and macro- and microscopic pathology. Tissue γ counting was performed to determine biodistribution of 131I-Hyp. RESULTS: 131I-Hyp based SPECT indicated clearly hot regions on ventricular walls which were all histologically proved as AMI. Complementally, the hot AMI regions on 131I-Hyp SPECT (infarct/myoc ratio of 15.3 ± 7.7) were inversely cold regions on 99mTc-sestamibi SPECT (infarct/myoc ratio of 0.029 ± 0.021). Autoradiography of heart slices showed 9.8 times higher 131I-Hyp uptake in infarcted over normal myocardium. With γ counting, the mean 131I-Hyp uptake in infarcts was 10.69 ID%/g, 12.05 times of that in viable myocardium. CONCLUSION: 131I-Hyp shows a potential for clinical detection of AMI once I-131 is substituted by its isotope like I-124 or I-123 for PET or SPECT, respectively.

A predictive model for first-trimester pregnancy inception after IVF-ET based on multimodal ultrasound evaluation of endometrial receptivity.

BACKGROUND: In-vitro fertilization-embryo transfer (IVF-ET) is a commonly used assisted reproductive technology. Its success depends on many factors, including endometrial receptivity. Endometrial receptivity can be evaluated by ultrasound, endometrial biopsy, and magnetic resonance imaging. Compared with the latter two methods, ultrasound has the advantages of wide availability, non-invasiveness, and low cost. Three-dimensional (3D) ultrasound imaging examines endometrial thickness, morphology, and blood vessels, which are associated with the success of embryo implantation. However, there are no reports of endometrial receptivity assessment by 3D ultrasound. Therefore, we aimed to evaluate endometrial receptivity using 3D ultrasound and construct a predictive model for first-trimester pregnancy inception following IVF-ET. METHODS: We performed a prospective observational study on infertile women who underwent IVF-ET between December 2019 and February 2021. These women had 3D ultrasound evaluations, measuring endometrial thickness, volume, pattern, morphology, peristalsis, uterine artery blood flow index, sub-endometrial blood flow index, and distribution pattern. We recorded the occurrence of first-trimester pregnancies in these women. Using Akaike information criterion (AIC) and backward stepwise regression, a first-trimester pregnancy prediction model was constructed based on the minimum AIC value and validated internally and externally. RESULTS: 111 women were enrolled, with 103 included in the analysis. Univariate and multiple logistic regression analyses showed that endometrial thickness and vascularization flow index (VFI) were independent factors associated with the occurrence of a pregnancy. The final prediction model corresponding to the minimum AIC value (65.166) was Y = - 6.131-0.182endometrial thickness + 0.542endometrial volume + 4.374VFI + 0.132age. In the test set, modeling cohort, and external validation cohort, the model showed satisfactory differentiation, with C index of 0.841 (95%CI 0.699-0.817), 0.727 (95%CI 0.619-0.815), and 0.745 (95%CI 0.671-0.840), respectively. The Hosmer-Lemeshow goodness of fit tests reported P = 0.865, 0.139, and 0.070, respectively, indicating a high agreement with the actual IVF-ET outcome. This model reached the highest diagnostic efficiency (sensitivity 88.9%, specificity 75%, Youden index 0.639) at a diagnostic cut-off value of ≥ 0.360. CONCLUSIONS: The predictive model based on endometrial receptivity evaluations by 3D ultrasound had high diagnostic efficiency and could be a simple and effective tool to predict first-trimester pregnancy inception after IVF-ET.

Construction of the amniotic fluid-derived exosomal ceRNA network associated with ventricular septal defect.

Ventricular septal defect (VSD) is the most frequent congenital cardiac malformations. Amniotic fluid (AF) contains a higher abundance of biological compounds that could reflect fetal health information. The aims of our study were to construct a competitive endogenous RNA (ceRNA) network based on AF-derived exosomal ncRNAs. We conducted whole transcriptome profiling in six pairs of AF-derived exosomes from VSD fetuses and matched healthy controls. A total of 1252 differentially expressed (DE) mRNAs, 256 DE-miRNAs and 1090 DE-lncRNAs were found to be significantly altered in the VSD group. We constructed a ceRNA regulatory network including 46 mRNAs, 11 miRNAs and 47 lncRNAs. The expression level of 6 hub RNAs were validated using qRT-PCR. In conclusion, AF-derived exosomal VSD-related ceRNAs provide a basis for a better understanding of the role of ncRNAs in the pathogenesis and mechanisms of VSD, which may lead to the discovery of potential diagnostic biomarkers for fetal VSD.

Combination of high-frequency ultrasound and virtual touch tissue imaging and quantification improve the diagnostic efficiency for mild carpal tunnel syndrome.

BACKGROUND: Carpal tunnel syndrome (CTS) is the most common entrapment symptom in the peripheral nerves. High-frequency ultrasound (HFUS) is widely used in the diagnosis of CTS. Virtual Touch Tissue Imaging and Quantification (VTIQ), which provides more information about the hardness of organization, is used to diagnose CTS. However, the data of diagnostic value of them in various degrees of CTS are limited. Whether the combination of HFUS and VTIQ can improve the diagnostic efficiency also remains unknown. The study aimed to explore the diagnostic value of HFUS and VTIQ in various degrees of CTS and whether combination of HFUS and VTIQ could improve the diagnostic efficiency of CTS. METHODS: A collection and analysis of 133 CTS patients and 35 volunteers from January 2016 to January 2019 were performed. We compared the clinical characteristics, cross-sectional area (CSA) value and shear wave velocity SWVmean value of CTS group with volunteer group. RESULTS: The CSA value and SWVmean value of CTS cohort were significantly higher than volunteer group (10.79 ± 2.88 vs. 8.06 ± 1.39, p < 0.001, 4.36 ± 0.95 vs. 3.38 ± 1.09, p < 0.001, respectively). The area under the curve (AUC) of receiver operating characteristic (ROC) curve of CSA value and SWVmean value were 0.794 and 0.757, respectively. Hierarchical analysis of CSA value and SWVmean value showed that the AUC in the moderate and severe CTS group were higher than in mild CTS group. Furthermore, the CSA value combined with SWVmean value used to diagnose mild CTS was 0.758, which was higher than that of single CSA value or single SWVmean value. CONCLUSIONS: Both HFUS and VTIQ technology were feasible to evaluate CTS. HFUS was suitable for use in diagnosis of moderate and severe CTS. For mild CTS, combination of HFUS and VTIQ was relevant to improve the diagnostic efficiency of CTS.

Proteomics insights into the effects of MSTN on muscle glucose and lipid metabolism in genetically edited cattle.

The molecular mechanism underlying myostatin (MSTN)-regulated metabolic cross-talk remains poorly understood. In this study, we performed comparative proteomic and phosphoproteomic analyses of gluteus muscle tissues from MSTN-/- transgenic cattle using a shotgun-based tandem mass tag (TMT) 6-plex labeling method to explore the signaling pathway of MSTN in metabolic cross-talk and cellular metabolism during muscle development. A total of 72 differentially expressed proteins (DEPs) and 36 differentially expressed phosphoproteins (DEPPs) were identified in MSTN-/- cattle compared to wild-type cattle. Bioinformatics analyses showed that MSTN knockout increased the activity of many key enzymes involved in fatty acid ß-oxidation and glycolysis processes in cattle. Furthermore, comprehensive pathway analyses and hypothesis-driven AMP-activated protein kinase (AMPK) activity assays suggested that MSTN knockout triggers the activation of AMPK signaling pathways to regulate glucose and lipid metabolism by increasing the AMP/ATP ratio. Our results shed new light on the potential regulatory mechanism of MSTN associated with metabolic cross-talk in muscle development, which can be used in animal breeding to improve meat production in livestock animals, and can also provide valuable insight into treatments for obesity and diabetes mellitus in humans.

Risk prediction for postpartum depression based on random forest. / 随机森林算法在产后抑郁风险预测中的应用.

OBJECTIVES: To explore the application of random forest algorithm in screening the risk factors and predictive values for postpartum depression. METHODS: We recruited the participants from a tertiary hospital between June 2017 and June 2018 in Changsha City, and followed up from pregnancy up to 4-6 weeks postpartum.Demographic economics, psychosocial, biological, obstetric, and other factors were assessed at first trimesters with self-designed obstetric information questionnaire and the Chinese version of Edinburgh Postnatal Depression Scale (EPDS). During 4-6 weeks after delivery, the Chinese version of EPDS was used to score depression and self-designed questionnaire to collect data of delivery and postpartum. The data of subjects were randomly divided into the training data set and the verification data set according to the ratio of 3꞉1. The training data set was used to establish the random forest model of postpartum depression, and the verification data set was used to verify the predictive effects via the accuracy, sensitivity, specificity, positive predictive value, negative predictive value, and AUC index. RESULTS: A total of 406 participants were in final analysis. Among them, 150 of whom had EPDS score ≥9, and the incidence of postpartum depression was 36.9%. The predictive effects of random forest model in the verification data set were at accuracy of 80.10%, sensitivity of 61.40%, specificity of 89.10%, positive predictive value of 73.00%, negative predictive value of 82.80%, and AUC index of 0.833. The top 10 predictive influential factors that screening by the variable importance measure in random forest model was antenatal depression, economic worries after delivery, work worries after delivery, free triiodothyronine in first trimesters, high-density lipoprotein in third trimester, venting temper to infants, total serum cholesterol and serum triglyceride in first trimester, hematocrit and serum triglyceride in third trimester. CONCLUSIONS: Random forest has a great advantage in risk prediction for postpartum depression. Through comprehensive evaluation mechanism, it can identify the important influential factors for postpartum depression from complex multi-factors and conduct quantitative analysis, which is of great significance to identify the key factors for postpartum depression and carry out timely and effective intervention.

Antitumor activity in colorectal cancer induced by hinokiflavone.

BACKGROUND AND AIM: Colorectal cancer is one of the most common malignant disease worldwide with highly metastatic potential. Identification of effective therapeutic treatment overcoming such disease is an urgent need. Our study focuses on hinokiflavone as an antitumor agent against colorectal cancer. METHODS: MTT assay, cell colony formation assay, Hoechst staining, flow cytometry, Western blot analysis, real-time polymerase chain reaction, and migration and invasion assay were performed to identify the effects of hinokiflavone on cell proliferation, apoptosis, and metastasis. CT26 tumor-bearing mice model was conducted to explore the antitumor activity of hinokiflavone in vivo. Immunohistochemistry staining was used to detect the protein expression of Ki-67, cleaved caspase-3, and MMP9 in treated tumors. Acute toxicity was evaluated by serological and hematological analyses, and drug side effect on organs was evaluated by hematoxylin and eosin staining. RESULTS: Hinokiflavone reduced the proliferation, migration, and invasion and promoted the apoptosis in colorectal tumor cells in vitro. Treatment of hinokiflavone at a tolerable and safe dose (50 mg/kg) significantly suppressed tumor growth in mice bearing CT26 tumors by reducing tumor proliferation and metastasis and inducing apoptosis. Mechanically, treatment of hinokiflavone induced apoptosis by loss of mitochondrial transmembrane potential and increased reactive oxygen species generation. CONCLUSIONS: Hinokiflavone suppressed colorectal tumor cell proliferation, induced apoptosis via the reactive oxygen species-mitochondria-mediated apoptotic pathway, and inhibited tumor cell migration and invasion. Antitumor activity of hinokiflavone was also validated in mice model without observed toxicity. Our findings suggested that the plant-derived hinokiflavone could be used as an antitumor agent against colorectal cancer.

Subcellular distribution and tolerance of cadmium in Canna indica L.

Canna indica L. is a promising species for heavy metal phytoremediation due to its fast growth rate and large biomass. However, few studies have investigated cadmium (Cd) tolerance mechanisms. In the present study, Canna plants were cultivated under hydroponic conditions with increasing Cd concentrations (0, 5, 10, 15 mg/L). We found that the plants performed well under 5 mg/L Cd2+ stress, but damage was observed under higher Cd exposure, such as leaf chlorosis, growth inhibition, a decreased chlorophyll content, and destruction of the ultrastructure of leaf cells. Additionally, Canna alleviated Cd toxicity to a certain extent. After Canna was exposed to 5, 10 and 15 mg/L Cd2+ for 45 d, the highest Cd concentration was exhibited in roots, which was almost 17-47 times the Cd concentration in leaves and 8-20 times that in stems. At the subcellular level, cellular debris and heat-stable proteins (HSPs) were the main binding sites for Cd, and the proportion of Cd in the two subcellular fractions accounted for 71.4-94.2% of the total Cd. Furthermore, we found that granules could participate in the detoxification process when Cd stress was enhanced. Our results indicated that Canna indica L. can tolerate Cd toxicity by sequestering heavy metals in root tissues, fencing out by cell wall, and binding with biologically detoxified fractions (granules and HSPs).

Knockdown of protein kinase CK2 blocked gene expression mediated by brain-derived neurotrophic factor-induced serum response element.

One of the principal signaling pathway outcomes from brain-derived neurotrophic factor (BDNF) is the activation of antiapoptotic pathways. In addition to the role of extracellular signal-regulated kinase 1/2 and phosphatidylinositol-3 kinase, BDNF activates protein kinase CK2 to mediate its neuroprotective effect. The inhibition of CK2 activity has been shown to induce apoptosis. Although serum response element (SRE)-mediated transcription has been reported to be activated by BDNF and that the phosphorylation of serum response factor (SRF) by CK2 has been shown to enhance its DNA binding activity, the biological relevance of these interactions remains largely unclear. In the present study, we found that SRE-mediated transcription, CK2 activity, and SRF phosphorylation increased in PC12 cells under BDNF treatment. The transfection of CK2α siRNA blocked the enhancing effect of BDNF on SRE-mediated transcription, SRF phosphorylation, and Mcl-1 gene expression. Moreover, the blockade of CK2 diminished the antiapoptotic effects of BDNF on SRE-mediated transcription, Mcl-1 gene expression, and cell viability under rotenone-induced cytotoxicity. Our data may assist in the development of therapeutic strategies for inhibiting apoptosis during neurodegeneration.

Human mitochondrial DNA haplogroup M8a influences the penetrance of m.8684C>T in Han Chinese men with non-obstructive azoospermia.

RESEARCH QUESTION: What is the role of mitochondrial DNA (mtDNA) in the pathogenesis of non-obstructive azoospermia (NOA)? DESIGN: mtDNA genome sequencing followed by an independent population validation were performed in 628 NOA cases and 584 healthy controls. Antioxidant capacity of serum was evaluated in 54 randomly selected cases out of 536 and 49 out of 489 controls. RESULTS: In the screening stage, 13 mtDNA haplogroups (hg) were ascertained, and 10 susceptible variants were observed. In the validation stage, hg M8* in individuals was found to be associated with increased risk of NOA [odds ratio (OR) 2.61, 95% confidence interval (CI) 1.47-4.61] (P=0.001). Unexpectedly, the frequency of m.8684C>T, the defining marker for hg M8a, was also higher in NOA (OR 4.14, 95% CI 1.56-11.03) (P=0.002). Subsequently, the frequency distributions were compared among the sub-hg of hg M8* (including hg M8a, C and Z) and, intriguingly, no significance was found in hg C and Z. Additionally, the level of total antioxidant capacity was significantly decreased (P<0.05) compared with the control group. CONCLUSIONS: hg M8a background in general played an active role in the penetrance of 8684C>T in NOA, and mtDNA genetic variants (causing low antioxidant levels) might increase mtDNA damage and impair normal spermatogenesis.

Magnetic Porous Molecularly Imprinted Polymers Based on Surface Precipitation Polymerization and Mesoporous SiO2 Layer as Sacrificial Support for Efficient and Selective Extraction and Determination of Chlorogenic Acid in Duzhong Brick Tea.

Magnetic porous molecularly imprinted polymers (MPMIPs) for rapid and efficient selective recognition of chlorogenic acid (CGA) were effectively prepared based on surface precipitation polymerization using CGA as template, 4-vinylpyridine (4-VP) as functional monomer, and mesoporous SiO2 (mSiO2) layer as sacrificial support. A computational simulation by evaluation of electronic binding energy is used to optimize the stoichiometric ratio between CGA and 4-VP (1:5), which reduced the duration of laboratory trials. The porous MIP shell and the rid of solid MIPs by magnet gave MPMIPs high binding capacity (42.22 mg/g) and fast kinetic binding (35 min). Adsorption behavior between CGA and MPMIPs followed Langmuir equation and pseudo-first-order reaction kinetics. Furthermore, the obtained MPMIPs as solid phase adsorbents coupled with high performance liquid chromatography (HPLC) were employed for selective extraction and determination of CGA (2.93 ± 0.11 mg/g) in Duzhong brick tea. The recoveries from 91.8% to 104.2%, and the limit of detection (LOD) at 0.8 µg/mL were obtained. The linear range (2.0⁻150.0 µg/mL) was wide with R² > 0.999. Overall, this study provided an efficient approach for fabrication of well-constructed MPMIPs for fast and selective recognition and determination of CGA from complex samples.

Comparative muscle proteomics/phosphoproteomics analysis provides new insight for the biosafety evaluation of fat-1 transgenic cattle.

The biosafety of fat-1 transgenic cattle has been a focus of our studies since the first fat-1 transgenic cow was born. In this study, we used tandem mass tag labeling, TiO2 enrichment, and nanoscale liquid chromatography coupled with tandem mass spectrometry (nanol LC-MS/MS) to compare proteomic and phosphoproteomic profiling analyses of muscle between fat-1 transgenic cows and wild-type cows. A total of 1555 proteins and 900 phosphorylation sites in 159 phosphoproteins were identified in the profiling assessments, but only four differentially expressed proteins and nine differentially expressed phosphopeptides were detected in fat-1 transgenic cows relative to wild-type cows. Bioinformatics analyses showed that all of the identified proteins and phosphoproteins were mainly related to the metabolic processes of three major nutrients: carbohydrates, lipids, and proteins. All of these differentially expressed proteins might take part in DNA recombination, repair, and regulation of the immune system. In conclusion, most of the identified proteins and phosphoproteins exhibited few changes. Our results provide new insights into the biosafety of fat-1 transgenic cattle.

Noninvasive evaluation of gastric emptying and gastric wall thickness in SLE patients.

OBJECTIVE: The objective of this study is to evaluate the gastric emptying in patients with systemic lupus erythematosus (SLE) with gastrointestinal involvement using three-dimensional (3D) ultrasonography. METHODS: The gastric emptying times at 25% (T1), 50% (T2), and 75% (T3) of SLE patients with gastrointestinal involvement (n = 40) and healthy controls (n = 80) were evaluated and compared. In addition, the correlations among the gastric wall thickness, SLE disease activity index (SLEDAI), and upper gastrointestinal symptoms were calculated. RESULTS: The gastric wall thickness was correlated with the SLEDAI (r = 0.928, p < 0.001) and the upper gastrointestinal symptom index (r = 0.848, p < 0.001). The emptying times T1, T2, and T3 of the SLE patients were 17.08 ± 2.65 min (mean ± standard deviation), 39.85 ± 6.54 min, and 83.58 ± 7.12 min, respectively. For healthy controls, they were 19.65 ± 5.39 min, 41.08 ± 7.51 min, and 70.34 ± 8.03 min. The T1 of the SLE patients was shorter (p < 0.01), while the T3 was longer (p < 0.001). Moreover, T3 in the SLE group had the best correlation with the upper gastrointestinal symptom index (r = 0.553, p < 0.001). T1 in the SLE group was anti-correlated with early satiety (r = -0.366, p < 0.05). CONCLUSIONS: Combining the emptying times T1 and T3, as well as the gastric wall thickness, the SLEDAI and the upper gastrointestinal symptoms index can provide accurate clinical diagnosis of SLE with gastric involvement.

Oncoprotein YAP regulates the spindle checkpoint activation in a mitotic phosphorylation-dependent manner through up-regulation of BubR1.

The transcriptional co-activator YAP (Yes-associated protein) functions as an oncogene; however, it is largely unclear how YAP exerts its oncogenic role. In this study, we further explored the functional significance of YAP and its mitotic phosphorylation in the spindle checkpoint. We found that the dynamic mitotic phosphorylation of YAP was CDC14-dependent. We also showed that YAP was required for the spindle checkpoint activation induced by spindle poisons. Mitotic phosphorylation of YAP was required for activation of the spindle checkpoint. Furthermore, enhanced expression of active YAP hyperactivated the spindle checkpoint and induced mitotic defects in a mitotic phosphorylation-dependent manner. Mechanistically, we documented that mitotic phosphorylation of YAP controlled transcription of genes associated with the spindle checkpoint. YAP constitutively associated with BubR1 (BUB1-related protein kinase), and knockdown of BubR1 relieved YAP-driven hyperactivation of the spindle checkpoint. Finally, we demonstrated that YAP promoted epithelial cell invasion via both mitotic phosphorylation and BubR1-dependent mechanisms. Together, our results reveal a novel link between YAP and the spindle checkpoint and indicate a potential mechanism underlying the oncogenic function of YAP through dysregulation of the spindle checkpoint.