The Beneficial Effects of Lactobacillus GG Therapy on Liver and Drinking Assessments in Patients with Moderate Alcohol-Associated Hepatitis.

INTRODUCTION: We investigated the effect of daily oral Lactobacillus rhamnosus GG (LGG) in reducing liver injury/severity and drinking in patients with alcohol use disorder and moderately severe alcohol-associated hepatitis. METHODS: Forty-six male and female individuals with alcohol use disorder and moderate alcohol-associated hepatitis (12 ≤ model for end-stage liver disease score < 20, aged 21-67 years) received either LGG (n = 24) or placebo (n = 22). Data were collected/assessed at baseline and at 1, 3, and 6 months. RESULTS: LGG treatment was associated with a significant reduction in liver injury after 1 month. Six months of LGG treatment reduced heavy drinking levels to social or abstinence levels. DISCUSSION: LGG treatment was associated with an improvement in both liver injury and drinking.

DNA-Binding One Finger Transcription Factor PhDof28 Regulates Petal Size in Petunia.

Petal size is a key indicator of the ornamental value of plants, such as Petunia hybrida L., which is a popular ornamental species worldwide. Our previous study identified a flower-specific expression pattern of a DNA-binding one finger (Dof)-type transcription factor (TF) PhDof28, in the semi-flowering and full-flowering stages of petunia. In this study, subcellular localization and activation assays showed that PhDof28 was localized in the cell nucleus and could undergo in vitro self-activation. The expression levels of PhDof28 tended to be significantly up-regulated at the top parts of petals during petunia flower opening. Transgenic petunia 'W115' and tobacco plants overexpressing PhDof28 showed similar larger petal phenotypes. The cell sizes at the middle and top parts of transgenic petunia petals were significantly increased, along with higher levels of endogenous indole-3-acetic acid (IAA) hormone. Interestingly, the expression levels of two TFs, PhNAC100 and PhBPEp, which were reported as negative regulators for flower development, were dramatically increased, while the accumulation of jasmonic acid (JA), which induces PhBPEp expression, was also significantly enhanced in the transgenic petals. These results indicated that PhDof28 overexpression could increase petal size by enhancing the synthesis of endogenous IAA in petunias. Moreover, a JA-related feedback regulation mechanism was potentially activated to prevent overgrowth of petals in transgenic plants. This study will not only enhance our knowledge of the Dof TF family, but also provide crucial genetic resources for future improvements of plant ornamental traits.

Preparation of Biomass Biochar with Components of Similar Proportions and Its Methylene Blue Adsorption.

Rapeseed straw, bagasse, and walnut peel have a large amount of resource reserves, but there are few technologies for high value-added utilization. In the research of biochar, walnut green husk is rarely used as raw material. In addition, the three main components of biomass (lignin, cellulose, and hemicellulose) are present in similar proportions, and the differences between the physical and chemical properties of biochar prepared with similar amounts of biomass raw materials are not clear. Using three kinds of biomass of the same quality as raw materials, biochar was prepared via pyrolysis at 400 °C, and activated carbon was prepared via CO2 activation at 800 °C. The results showed that the pore numbers of the three kinds of biochar increased after activation, resulting in the increase of the specific surface area. The resulting numbers were 352.99 m2/g for sugarcane bagasse biochar (SBB)-CO2, 215.04 m2/g for rapeseed straw biochar (RSB)-CO2, and 15.53 m2/g for walnut green husk biochar (WGB)-CO2. Ash increased the amount of carbon formation, but a large amount of ash caused biochar to form a perforated structure and decreased the specific surface area (e.g., WGB), which affected adsorption ability. When the three main components were present in similar proportions, a high content of cellulose and lignin was beneficial to the preparation of biochar. The adsorption value of MB by biochar decreased with the increase of biomass ash content. After activation, the maximum adsorption value of MB for bagasse biochar was 178.17 mg/g, rapeseed straw biochar was 119.25 mg/g, and walnut peel biochar was 85.92 mg/g when the concentration of methene blue solution was 300 mg/L and the biochar input was 0.1 g/100 mL at room temperature. The adsorption of MB by biochar in solution occurs simultaneously with physical adsorption and chemical adsorption, with chemical adsorption being dominant. The optimal MB adsorption by SBB-CO2 was dominated by multimolecular-layer adsorption. This experiment provides a theoretical basis for the preparation of biochar and research on its applications in the future.

The C2H2-type zinc finger protein PhZFP1 regulates cold stress tolerance by modulating galactinol synthesis in Petunia hybrida.

The C2H2 zinc finger proteins (ZFPs) play essential roles in regulating cold stress responses. Similarly, raffinose accumulation contributes to freezing stress tolerance. However, the relationship between C2H2 functions and raffinose synthesis in cold tolerance remains uncertain. Here, we report the characterization of the cold-induced C2H2-type zinc finger protein PhZFP1 in Petunia hybrida. PhZFP1 was found to be predominantly localized in the nucleus. Overexpression of PhZFP1 conferred enhanced cold tolerance in transgenic petunia lines. In contrast, RNAi mediated suppression of PhZFP1 led to increased cold susceptibility. PhZFP1 regulated the expression of a range of abiotic stress responsive-genes including genes encoding proteins involved in reactive oxygen species (ROS) scavenging and raffinose metabolism. The accumulation of galactinol and raffinose, and the levels of PhGolS1-1 transcripts, were significantly increased in PhZFP1-overexpressing plants and decreased in PhZFP1-RNAi plants under cold stress. Moreover, the galactinol synthase (GolS)-encoding gene PhGolS1-1 was identified as a direct target of PhZFP1. Taken together, these results demonstrate that PhZFP1 functions in cold stress tolerance by modulation of galactinol synthesis via regulation of PhGolS1-1. This study also provides new insights into the mechanisms underlying C2H2 zinc finger protein-mediated cold stress tolerance, and has identified a candidate gene for improving cold stress tolerance.

Analysis of alcohol use, consumption of micronutrient and macronutrients, and liver health in the 2017-2018 National Health and Nutrition Examination Survey.

BACKGROUND: Alcohol use is a major global healthcare burden that contributes to numerous adverse health outcomes, including liver disease. Many factors influence individual susceptibility to alcohol-associated diseases, including nutritional factors. The objective of the current study was to examine inter-relations among alcohol, dietary micronutrients and macronutrient consumption, and liver health by analyzing data from the 2017-2018 National Health and Nutrition Examination Survey (NHANES). METHODS: Based on self-reported alcohol consumption, NHANES respondents were assigned to one of four categories: never drinkers (lifetime abstainers), non-drinkers (past-year abstainers), moderate drinkers (1/2 drinks per day for females/males, respectively), and heavy drinkers (>1/>2 drinks per day for females/males, respectively, and/or frequent binge drinking). Survey-weighted regression analyses (adjusted for gender, age, race, education, and body mass index) were performed to examine associations between alcohol intake, dietary, and liver health characteristics. RESULTS: Individuals categorized as heavy drinkers were significantly younger, most often well-educated males with low incidences of diabetes and other comorbidities. They consumed the most overall calories and various micronutrients, indicating a diet that was not necessarily nutrient poor. Neither moderate nor heavy drinkers had liver steatosis or fibrosis as measured by liver elastography, although heavy drinkers had modestly elevated plasma biomarkers of liver injury, including ALT, AST, and GGT, compared with the other groups. CONCLUSIONS: Our findings suggest that the category of heavy drinkers in the 2017-2018 NHANES consisted of generally healthy individuals with high-energy intake and no evidence of liver steatosis or fibrosis. However, slightly increased plasma liver markers may indicate a risk of future progression to more advanced stages of liver disease over time in some individuals. Several limitations should be considered when interpreting these data, including the potential misclassification of drinking categories and the lack of standardized cutoff scores for fatty liver as assessed by elastography, among others.

TGF-ß1/SMOC2/AKT and ERK axis regulates proliferation, migration, and fibroblast to myofibroblast transformation in lung fibroblast, contributing with the asthma progression.

BACKGROUND: Asthma is a common chronic respiratory disease that influences 300 million people all over the world. However, the pathogenesis of asthma has not been fully elucidated. It has been reported that transforming growth factor-ß (TGF-ß) can activate myofibroblasts. Moreover, the fibroblast to myofibroblast transformation (FMT) can be triggered by TGF-ß, which is a major mediator of subepithelial fibrosis. Secreted modular calcium-binding protein 2 (SMOC2) is a member of cysteine (SPARC) family and is involved in the progression of multiple diseases. However, its role in asthma remains poorly understood. RT-qPCR evaluated the expression of SMOC2. Bromodeoxyuridine assay and wound-healing assay detected the proliferation and migration of lung fibroblasts, respectively. IF staining was performed to assess the expression of α-smooth muscle actin (α-SMA). Western blot analysis detected the levels of proteins. Flow cytometry was utilized for determination of the number of myofibroblasts. RESULTS: We found the expression of SMOC2 was upregulated by the treatment of TGF-ß1 in lung fibroblasts. In addition, SMOC2 promoted the proliferation and migration of lung fibroblasts. More importantly, SMOC2 accelerated FMT of lung fibroblasts. Furthermore, SMOC2 was verified to control the activation of AKT and ERK. Rescue assays showed that the inhibition of AKT and ERK pathway reversed the promoting effect of SMOC2 overexpression on proliferation, migration and FMT in lung fibroblasts. CONCLUSIONS: This work demonstrated that SMOC2 modulated TGF-ß1-induced proliferation, migration and FMT in lung fibroblasts and may promote asthma, which potentially provided a novel therapeutic target for the management of asthma.

Transcriptomic and GC-MS Metabolomic Analyses Reveal the Sink Strength Changes during Petunia Anther Development.

Petunia, which has been prevalently cultivated in landscaping, is a dicotyledonous herbaceous flower of high ornamental value. Annually, there is a massive worldwide market demand for petunia seeds. The normal development of anther is the necessary prerequisite for the plants to generate seeds. However, the knowledge of petunia anther development processes is still limited. To better understand the mechanisms of petunia anther development, the transcriptomes and metabolomes of petunia anthers at three typical development stages were constructed and then used to detect the gene expression patterns and primary metabolite profiles during the anther development processes. Results suggested that there were many differentially-expressed genes (DEGs) that mainly participated in photosynthesis and starch and sucrose metabolism when DEGs were compared between the different development stages of anthers. In this study, fructose and glucose, which were involved in starch and sucrose metabolism, were taken as the most important metabolites by partial least-squares discriminate analysis (PLS-DA). Additionally, the qRT-PCR analysis of the photosynthetic-related genes all showed decreased expression trends along with the anther development. These pieces of evidence indicated that the activities of energy and carbohydrate metabolic pathways were gradually reduced during all the development stages of anther, which affects the sink strength. Overall, this work provides a novel and comprehensive understanding of the metabolic processes in petunia anthers.

An anther-specific gene PhGRP is regulated by PhMYC2 and causes male sterility when overexpressed in petunia anthers.

KEY MESSAGE: An anther-specific GRP gene, regulated by PhMYC2 , causes a significant reduction of male fertility when overexpressed in petunia, and its promoter is efficient in genetic engineering of male-sterile lines. Glycine-rich proteins (GRPs) play important roles in plant anther development; however, the underlying mechanisms and related regulatory networks are poorly understood. In this study, a novel glycine-rich family gene designated as PhGRP was isolated from Petunia hybrida 'Fantasy Red'. The qRT-PCR analysis showed that it expressed specifically in anthers, and its expression peaked earlier than those well-known tapetum-specific genes, such as TA29, and several genes with the classic cis-regulatory element 'anther-box' in petunia during its anther development. The male fertility was significantly reduced in PhGRP overexpression lines, due to the abnormal formation of pollen wall. The PhGRP promoter (pPhGRP) could drive the GUS genes expressing specifically in the anthers of the transgenic Arabidopsis plants, indicating that the anther-specific characteristic of this promoter was conserved. In addition, when pPhGRP was used to drive the expression of BARNASE, complete male-sterile petunia lines were created without changes in vegetative organs and floral parts other than anthers. Finally, when pPhGRP was used as the bait to screen a yeast-one-hybrid (Y1H) library, a transcription factor (PhMYC2) belonging to the bHLH family was successfully selected, and the binding between pPhGRP and PhMYC2 was validated both by Y1H and dual-luciferase reporter assay. Overall, these results suggest that PhGRP, which is a male fertility-related gene that expresses specifically in anthers, is regulated by PhMYC2 and whose promoter can be used as an effective tool in the creation of male-sterile lines.

[Cellphone electromagnetic radiation damages the testicular ultrastructure of male rats].

OBJECTIVE: To investigate the influence of cellphone electromagnetic radiation (CER) on the testicular ultrastructure and the apoptosis of spermatogenic cells in male rats.atability, feasibility, applicability, and controllability in the construction of experimental animal models, we compared the major anatomic features of the penis of 20 adult beagle dogs with those of 10 adult men. Using microsurgical techniques, we performed cross-transplantation of the penis in the 20 (10 pairs) beagle dogs and observed the survival rate of the transplanted penises by FK506+MMF+MP immune induction. We compared the relevant indexes with those of the 10 cases of microsurgical replantation of the amputated penis. METHODS: Thirty adult male SD rats were equally randomized into a 2 h CER, a 4 h CER, and a normal control group, the former two groups exposed to 30 days of 900 MHz CER for 2 and 4 hours a day, respectively, while the latter left untreated. Then the changes in the ultrastructure of the testis tissue were observed under the transmission electron microscope and the apoptosis of the spermatogenic cells was determined by TUNEL. RESULTS: Compared with the normal controls, the rats of the 2 h CER group showed swollen basement membrane of seminiferous tubules, separated tight junction of Sertoli cells, increased cell intervals, apparent vacuoles and medullization in some mitochondria, and increased apoptosis of spermatogenic cells, mainly the apoptosis of primary spermatocytes (P<0.05 ). In comparison with the 2 h CER group, the animals of the 4 h CER group exhibited swollen basement membrane of seminiferous tubules, more separated tight junction of Sertoli cells, wider cell intervals, incomplete membrane of spermatogonial cells, fragments of cytoplasm, nuclear pyknosis and notch, slight dilation of perinuclear space, abnormalities of intracellular mitochondria with vacuoles, fuzzy structure, and fusion or disappearance of some cristae, and increased damage of mitochondria and apoptosis of spermatogenic cells, including the apoptosis of spermatogonial cells, primary spermatocytes, and secondary spermatocytes (P<0.05 ). CONCLUSIONS: CER can damage the testicular ultrastructure and increase the apoptosis of spermatogenic cells of the male rat in a time-dependent manner, and the apoptosis of spermatogenic cells may be associated with the damage to mitochondria.

Risk factors and related miRNA phenotypes of chronic pain after thoracoscopic surgery in lung adenocarcinoma patients.

Chronic postsurgical pain may have a substantial impact on patient's quality of life, and has highly heterogenous presentation amongst sufferers. We aimed to explore the risk factors relating to chronic pain and the related miRNA phenotypes in patients with lung adenocarcinoma after video-assisted thoracoscopic lobectomy to identify potential biomarkers. Our prospective study involved a total of 289 patients with early invasive adenocarcinoma undergoing thoracoscopic lobotomy and a follow-up period of 3 months after surgery. Blood was collected the day before surgery for miRNA detection and patient information including operation duration, duration of continuous drainage of the chest, leukocyte count before and after operation, and postoperative pain scores were recorded. Using clinical and biochemical information for each patient, the risk factors for chronic postsurgical pain and related miRNA phenotypes were screened. We found that chronic postsurgical pain was associated with higher body mass index; greater preoperative history of chronic pain; longer postoperative drainage tube retention duration; higher numerical rating scale scores one, two, and three days after surgery; and changes in miRNA expression, namely lower expression of miRNA 146a-3p and higher expression of miRNA 550a-3p and miRNA 3613-3p in peripheral blood (p < 0.05). Of these factors, patient body mass index, preoperative history of chronic pain, average numerical rating scale score after operation, and preoperative peripheral blood miRNA 550a-3P expression were independent risk factors for the development of chronic postsurgical pain. Identification of individual risk markers may aid the development and selection of appropriate preventive and control measures.

ALT poorly predicts Nonalcoholic Fatty Liver Disease (NAFLD) and liver fibrosis as determined by vibration-controlled transient elastography in adult National Health and Nutrition Examination Survey 2017-2018.

BACKGROUND: Non-alcoholic fatty liver disease is a growing problem in the United States, contributing to a range of liver disease as well as cardiovascular disease. ALT is the most widely used liver chemistry for NAFLD evaluation. We hypothesized that the normal range many laboratories use was too high, missing many patients with clinically important steatosis and/or fibrosis. METHODS: This study utilized 2017-2018 NHANES data including 9254 participants. We compared four different upper limits of normal for ALT with specific measurements of steatosis and liver stiffness as determined by liver elastography with FibroScan®. Liver stiffness was further characterized as showing any fibrosis or advanced fibrosis. After exclusions, our final pool was 4184 for liver stiffness measurement and 4183 for steatosis grade as measured by Controlled Attenuation Parameter (CAP). Using these variables, we performed logistic regression between ALT and CAP, and ALT and fibrosis/advanced fibrosis, and did a Receiver Operating Characteristic curve. RESULTS: Based on three of the most widely used cut off values for ALT, we found that ALT does not reliably rule out NAFLD in over 50% of cases. It also missed 45.9-64.2% of patients with liver fibrosis. CONCLUSIONS: Our study revealed that ALT is an inaccurate marker for NAFLD as measured by FibroScan® with CAP greater than or equal to 300 dB/m. Accuracy improved specific risk factors were considered. These data also showed that ALT was a poor marker for liver fibrosis. We conclude that there is no single ALT level that accurately predicts hepatic steatosis or fibrosis.

Interference fading suppression with fault-tolerant Kalman filter in phase-sensitive OTDR.

A multi-sensor information fusion algorithm based on fault-tolerant Kalman filter is proposed in phase-sensitive optical time-domain reflectometer (Φ-OTDR) system, for achieving fading-free distributed vibration sensing. Firstly, a fault-tolerant dual-core complementary array model is designed. The Rayleigh scattering signal denoising, and vibration existence judgment of localization points are carried out to obtain the differentiated frequency demodulation results of the sensing points of the dual-core fiber array. Then a fault-tolerant control strategy is used to determine the sensor weight coefficients and vibration judgment coefficients during data fusion processing, and array data fusion is carried out based on time series data using Kalman filter to realize error value identification and filling. The advantage of this method is the combination of redundant data in a complementary way to improve the system stability. The frequency response ranges from 10 Hz to 2400 Hz and the localization accuracy is 98.33%. The influence of key parameters on the frequency demodulation performance of fault-tolerant Kalman filter is discussed, and a standard deviation of 14.6 Hz and an average error of 7.6 Hz are obtained. The demodulation frequency data matrix obtained by the classical demodulation method has a demodulation error probability of 89.18%, which proves the widespread existence of demodulation errors in vibration signals. The fusion error of demodulation frequency is reduced to 0.25 Hz, the frequency demodulation accuracy reaches 100%, and the demodulation error caused by interference attenuation can be completely eliminated. This system based on fault-tolerant Kalman filter has the characteristics of simple multiplexing structure, interference fading resistance and stable demodulation performance.

One-month assessment of Th-cell axis related inflammatory cytokines, IL-17 and IL-22 and their role in alcohol-associated liver disease.

Introduction: Changes in the expression of cyto- and chemokines due to alcohol-associated liver disease (ALD) have been reported to be both protective and pathogenic. This study examined plasma levels of two key cytokines, Il-17 and Il-22, which construct the proinflammatory vs. anti-inflammatory axes across the spectrum of alcohol use disorder (AUD) and ALD including alcohol-associated hepatitis (AH) to determine the underlying status of the inflammation. Methods: Forty-two males and females aged 25-63 yrs. were grouped as healthy controls (HV[n=8]), AUD with no liver injury (AUDNLI [n=8]), AUD with liver injury (AUDLI [n=8]), non-severe alcohol-associated hepatitis (NSAH [n=9]), and severe alcohol-associated hepatitis (SAH [n=9]). Demographic, drinking, and clinical data were collected. Blood samples were collected at baseline (BL, all subjects) and during week 4 (W4, only patients) for IL-17 and IL-22; and statistically analyzed. Results: IL-17 was highly elevated in the SAH group both at BL and post-SOC. LTDH and BL IL-22 in non-severe AH patients were associated significantly. LTDH significantly predicted W4 IL-22 levels, positively (increasing) in NSAH and inversely (lowering) in SAH patients. BL and W4 IL-22 levels were significantly higher (4-fold, p≤0.001) in all AH patients compared to all AUD patients (AUROC=0.988, p≤0.001). IL-22 showed significant affinity with AST, AST: ALT ratio, total bilirubin, INR, and PT both at BL and W4. IL-22 was inversely associated with IL-1ß; and positively with TNF-α and IL-8 both at BL, and W4. BL IL-17 showed a positive correlation with MELD (p=0.017) in all AH patients. In SAH, > 2-fold W4 IL-17 level compared to BL showed significant within subjects' effects, p=0.006. In AUD patients without AH, the drop in IL-17 at W4 vs. BL showed a significant within subjects' effect, p=0.031. Discussion: Drinking chronicity predicted opposite effects in IL-22 levels in NSAH (antiinflammatory) and SAH (pro-inflammatory) patients at post-SOC. BL IL-22 levels differentiated AH patients robustly from the AUD patients (with or without liver injury); and showed corresponding increases stepwise with the stages of ALD. IL-22 was closely associated with progression and injury markers of the liver; and response to the cytokines of pro-inflammatory nature. Pro-inflammatory indicator of IL-17 cell axis, IL-17 showed a strong positive association with MELD, a severity indicator of AH.

Novel paradigms for the gut-brain axis during alcohol withdrawal, withdrawal-associated depression, and craving in patients with alcohol use disorder.

Introduction: Patients with alcohol use disorder (AUD) exhibit symptoms such as alcohol withdrawal, depression, and cravings. The gut-immune response may play a significant role in manifesting these specific symptoms associated with AUD. This study examined the role of gut dysfunction, proinflammatory cytokines, and hormones in characterizing AUD symptoms. Methods: Forty-eight AUD patients [men (n = 34) and women (n = 14)] aged 23-63 years were grouped using the Clinical Institute Withdrawal Assessment of Alcohol Scale (CIWA) as clinically significant (CS-CIWA [score > 10] [n = 22]) and a clinically not-significant group (NCS-CIWA [score ≤ 10] [n = 26]). Clinical data (CIWA, 90-day timeline followback [TLFB90], and lifetime drinking history [LTDH]) and blood samples (for testing proinflammatory cytokines, hormones, and markers of intestinal permeability) were analyzed. A subset of 16 AUD patients was assessed upon admission for their craving tendencies related to drug-seeking behavior using the Penn-Alcohol Craving Score (PACS). Results: CS-CIWA group patients exhibited unique and significantly higher levels of adiponectin and interleukin (IL)-6 compared to NCS-CIWA. In the CS group, there were significant and high effects of association for the withdrawal score with gut-immune markers (lipopolysaccharide [LPS], adiponectin, IL-6, and IL-8) and for withdrawal-associated depression with gut-immune markers (scored using MADRS with LPS, soluble cells of differentiation type 14 [sCD14], IL-6, and IL-8). Craving (assessed by PACS, the Penn-Alcohol Craving Scale) was significantly characterized by what could be described as gut dysregulation (LBP [lipopolysaccharide binding protein] and leptin) and candidate proinflammatory (IL-1ß and TNF-α) markers. Such a pathway model describes the heavy drinking phenotype, HDD90 (heavy drinking days past 90 days), with even higher effects (R2 = 0.955, p = 0.006) in the AUD patients, who had higher ratings for cravings (PACS > 5). Discussion: The interaction of gut dysfunction cytokines involved in both inflammation and mediating activity constitutes a novel pathophysiological gut-brain axis for withdrawal symptoms and withdrawal-associated depression and craving symptoms in AUD. AUD patients with reported cravings show a significant characterization of the gut-brain axis response to heavy drinking. Trial registration: ClinicalTrials.gov, identifier: NCT# 00106106.

SREBP2 promotes the viability, proliferation, and migration and inhibits apoptosis in TGF-ß1-induced airway smooth muscle cells by regulating TLR2/NF-κB/NFATc1/ABCA1 regulatory network.

Asthma is a respiratory disease with complex pathogenesis. Sterol-responsive element-binding proteins 2 (SREBP2) was found to bind to promoter sequences of ABCA1 to suppress ABCA1 promoter activity. This study aimed to explore the expression level of SREBP2 and ATP-binding cassette transporter A1 (ABCA1), and their effects on the development of airway smooth muscle cells (ASMCs) in asthma. ASMCs were treated with different concentrations of TGF-ß1 (0, 0.5, 1, 5 and 10 ng/mL). Short hairpin SREBP2 (shSREBP2), SREBP2, shABCA1 or ABCA1 were transfected into ASMCs. Cell viability, proliferation, apoptosis, migration, and the expression of SREBP2, ABCA1 and related pathway proteins were detected by MTT assay, Brdu staining, flow cytometer, Transwell assay, qRT-PCR, and Western blotting, respectively. The results showed that TGF-ß1 increased the viability, proliferation, migration and inhibited apoptosis in ASMCs. Moreover, TGF-ß1 also decreased the expression of ABCA1, cleaved caspase-3, cleaved PARP, E-cadherin, and increased the expression of vimentin, TLR2, p-p65 and NFATc1. SREBP2 knockdown alleviated these TGF-ß1-induced changes. SREBP2 overexpression inhibited ABCA1 expression and apoptosis, and promoted cell migration and the expression of TLR2, p-p65, NFATc1 in ASMCs. ABCA1 overexpression alleviated these SREBP2-induced promoting and inhibition effects. In conclusion, SREBP2 activates TLR2/NF-κB/NFATc1 regulatory network and promotes TGF-ß1-induced cell movement through inhibiting ABCA1 expression.

Prevalence and Treatment for Alcohol Use Disorders Based on Kentucky Medicaid 2012-2019 Datasets.

Alcohol use is the leading substance use in the United States. Persons with alcohol use disorder (AUD) face enormous health consequences and family problems. Analysis of Medicaid enrollee data is critical to understand different aspects of AUD and the treatment utilization for patients with AUD. Yearly patient-level data were constructed from the Kentucky 2012-2019 Medicaid claims data. ICD-9-CM and ICD-10-CM codes were used to identify patients with AUD and their comorbid conditions, the 11-digit National Drug Codes were used to identify medication treatments, and procedure codes were used to identify psychosocial and behavioral therapies. Logistic regression models were used to examine factors that were associated with AUD prevalence and AUD treatments. The prevalence of AUD trended up over time. Patients living in metro areas, between ages 45-54, having mental disorders, tobacco use, and with a family history of alcoholism had significantly higher rates of AUD. About 60% of patients diagnosed with AUD had major depressive disorder or anxiety. The treatment utilization for AUD also trended up from 2012 to 2019; however, it was still lower than 25% in 2019. Pharmacological treatments were used in only 2.89% of AUD cases in 2012, which increased to 8.13% in 2019. Psychosocial treatments were used in only 1.59% of AUD cases in 2012 that increased to 18.95% in 2019. The prevalence of AUD trended up over years. However, the treatment utilization for AUD was lower than 25%, even as of 2019. There is an urgent need for comprehensive, evidence-based, personalized AUD treatments.

Prevalence and Treatment Utilization of Patients Diagnosed with Depression and Anxiety Disorders Based on Kentucky Medicaid 2012-2019 Datasets.

Objectives: To examine the prevalence and treatment utilization of patients diagnosed with Depression and Anxiety Disorders (DAD) based on Kentucky Medicaid 2012-2019 datasets. Methods: The study was based on Kentucky Medicaid claims data from 2012 through 2019 for patients 14 years and older. We constructed yearly patient-level databases using ICD_9 CM and ICD_10 CM codes to identify the patients with DAD, using the Current Procedure Terminology (CPT) codes to identify individual psychotherapy and group psychotherapy and using the National drug codes to categorize pharmacotherapy. Based on these data, we constructed summary tables that reflected the trends in prevalence of DAD across eight Kentucky Medicaid regions and for different demographic subgroups. Next, we implemented logistic regression on the constructed yearly patient-level data to formally assess the impact of risk factors and treatments on the prevalence of DAD. The potential risk factors included age, gender, race/ethnicity, geographic characteristics, comorbidities such as alcohol use disorder and tobacco use. Results: The prevalence of DAD increased from 30.84% in 2012 to 36.04% in 2019. The prevalence of DAD was significantly higher in patients with the following characteristics: non-Hispanic white, females, aged between 45 and 54 years old, living in rural areas, having alcohol use disorder, and using tobaccos. Other than 2013, the utilization of pharmacotherapy maintained at about 62%. The utilization of psychotherapy increased over years from 24.4% in 2012 to 36.5% in 2019. Overall, the utilization of any treatment slightly increased from 70.9% in 2012 to 73.3% in 2019 except a drastic decline in 2013 due to the reduction of benzodiazepine prescription. Patients being whites, females, and living in rural areas were more likely to use pharmacotherapy, and patients living in rural areas were less likely to use psychotherapy than those residing in urban areas. Conclusion: The prevalence of DAD has increased over time from 2012 to 2019. The utilization of pharmacotherapy maintained at 62% over eight years except 2013, and the utilization of psychotherapy has steadily increased over time.

Illustration of Gut-Thyroid Axis in Alcohol Use Disorder: Interplay of Gut Dysfunction, Pro-Inflammatory Responses, and Thyroid Function.

(1) Background: Heavy and chronic alcohol drinking leads to altered gut dysfunction, coupled with a pro-inflammatory state. Thyroid-associated hormones and proteins may be dysregulated by heavy and chronic alcohol intake; however, the mechanism for altered gut-derived changes in thyroid function has not been studied thus far. This study investigates the role of alcohol-induced gut dysfunction and pro-inflammatory cytokine profile in the thyroid function of patients with alcohol use disorder (AUD). (2) Methods: Male and female AUD patients (n = 44) were divided into Gr.1, patients with normal thyroid-stimulating hormone (TSH) levels (n = 28, 0.8 ≤ TSH ≤ 3 mIU/L); and Gr.2, patients with clinically elevated TSH levels (n = 16, TSH > 3 mIU/L). Demographics, drinking measures, comprehensive metabolic panels, and candidate thyroid markers (TSH, circulating triiodothyronine (T3), and free thyroxine (fT4)) were analyzed. Gut-dysfunction-associated markers (lipopolysaccharide (LPS), LPS-binding protein (LBP), and soluble LPS-induced pathogen-associated protein (sCD14)), and candidate pro-inflammatory cytokines (IL-1ß, TNF-α, IL-6, IL-8, MCP-1, PAI-1) were also evaluated. (3) Results: Patients in both groups presented with a borderline overweight BMI category. Gr.2 reported numerically higher indices of chronic and heavy drinking patterns than Gr.1. The fT4 levels were elevated, while T3 was within normal limits in both groups. The gut dysfunction markers LBP and sCD14 were numerically elevated in Gr.2 vs. Gr.1, suggesting subtle ongoing changes. Candidate pro-inflammatory cytokines were significantly elevated in Gr.2, including IL-1 ß, MCP-1, and PAI-1. Gr.2 showed a strong and statistically significant effect on the gut-immune-thyroid response (r = 0.896, 36 p = 0.002) on TSH levels in a multivariate regression model with LBP, sCD14, and PAI-1 levels as upstream variables in the gut-thyroid pathway. In addition, AUROC analysis demonstrated that many of the cytokines strongly predicted TSH in Gr.2, including IL-6 (area = 0.774, 39 p < 0.001) and TNF-α (area = 0.708, p = 0.017), among others. This was not observed in Gr.1. Gr.2 demonstrated elevated fT4, as well as TSH, which suggests that there was subclinical thyroiditis with underlying CNS dysfunction and a lack of a negative feedback loop. (4) Conclusions: These findings reveal the toxic effects of heavy and chronic drinking that play a pathological role in thyroid gland dysregulation by employing the gut-brain axis. These results also emphasize potential directions to carefully evaluate thyroid dysregulation in the overall medical management of AUD.

Genome-Wide Identification and Characterization of JAZ Protein Family in Two Petunia Progenitors.

Jasmonate ZIM-domain (JAZ) family proteins are the key repressors in the jasmonate signaling pathway and play crucial roles in plant development, defenses, and responses to stresses. However, our knowledge about the JAZ protein family in petunia is limited. This research respectively identified 12 and 16 JAZ proteins in two Petunia progenitors, Petunia axillaris and Petunia inflata. Phylogenetic analysis showed that the 28 proteins could be divided into four groups (Groups A-D) and further classified into six subgroups (A1, A2, B1, B3, C, and D1); members in the same subgroup shared some similarities in motif composition and sequence structure. The Ka/Ks ratios of seven paralogous pairs were less than one, suggesting the petunia JAZ family might have principally undergone purifying selection. Quantitative real-time PCR (qRT-PCR) analysis revealed that PaJAZ genes presented differential expression patterns during the development of flower bud and anther in petunia, and the expression of PaJAZ5, 9, 12 genes was generally up-regulated after MeJA treatment. Subcellular localization assays demonstrated that proteins PaJAZ5, 9, 12 were localized in nucleus. Yeast two hybrid (Y2H) elucidated most PaJAZ proteins (PaJAZ1-7, 9, 12) might interact with transcription factor MYC2. This study provides insights for further investigation of functional analysis in petunia JAZ family proteins.

Integrating Transcriptomic and GC-MS Metabolomic Analysis to Characterize Color and Aroma Formation during Tepal Development in Lycoris longituba.

Lycoris longituba, belonging to the Amaryllidaceae family, is a perennial bulb bearing flowers with diverse colors and fragrance. Selection of cultivars with excellent colored and scented flowers has always been the breeding aim for ornamental plants. However, the molecular mechanisms underlying color fading and aroma production during flower expansion in L. longituba remain unclear. Therefore, to systematically investigate these important biological phenomena, the tepals of L. longituba from different developmental stages were used to screen and analyze the metabolic components and relevant genes. Utilizing the Illumina platform, a total of 144,922 unigenes were obtained from the RNA-Seq libraries. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis indicated that the phenylpropanoid biosynthesis and flavonoid biosynthesis pathways might play important roles during color and aroma changes. Metabolomic analysis identified 29 volatile organic components (VOCs) from different developmental stages of L. longituba tepals, and orthogonal partial least-squares discriminate analysis (OPLS-DA) revealed that trans-ß-ocimene-a terpene-was the most important aroma compound. Meanwhile, we found the content of anthocyanin was significantly reduced during the tepal color fading process. Then, we identified two dihydroflavonol-4-reductase (DFR) and three terpene synthase (TPS) genes, for which expression changes coincided with the production patterns of anthocyanins and trans-ß-ocimene, respectively. Furthermore, a number of MYB and bHLH transcription factors (TFs) which might be involved in color- and aroma-formation were also identified in L. longituba tepal transcriptomes. Taken together, this is the first comprehensive report of the color and fragrance in tepals of L. longituba and these results could be helpful in understanding these characteristics and their regulation networks.