Thermally Robust Solvent-Free Liquid Polyplexes for Heat-Shock Protection and Long-Term Room Temperature Storage of Therapeutic Nucleic Acids.

Nucleic acid therapeutics have attracted recent attention as promising preventative solutions for a broad range of diseases. Nonviral delivery vectors, such as cationic polymers, improve the cellular uptake of nucleic acids without suffering the drawbacks of viral delivery vectors. However, these delivery systems are faced with a major challenge for worldwide deployment, as their poor thermal stability elicits the need for cold chain transportation. Here, we demonstrate a biomaterial strategy to drastically improve the thermal stability of DNA polyplexes. Importantly, we demonstrate long-term room temperature storage with a transfection efficiency maintained for at least 9 months. Additionally, extreme heat shock studies show retained luciferase expression after heat treatment at 70 °C. We therefore provide a proof of concept for a platform biotechnology that could provide long-term room temperature storage for temperature-sensitive nucleic acid therapeutics, eliminating the need for the cold chain, which in turn would reduce the cost of distributing life-saving therapeutics worldwide.

A comparative analysis of the levels and drivers of regional coordinated development in the Yangtze River Economic Belt and Yellow River Basin, China.

Positioned in the era of the transformation of China's primary social contradictions, this study delves into the new connotations of regional coordinated development(RCD) from the perspective of "factors" coordination within the region and constructs an RCD evaluation system from five subsystems of regional economic coordination(REC), urban-rural coordination(URC), economic and social coordination(EASC), resource and environmental coordination(RAEC), and material and spiritual civilization coordination(MASCC). Then, the Entropy weight-TOPSIS model is used to evaluate the RCD levels of the 19 provinces located in the Yangtze River Economic Belt(YREB) and Yellow River Basin(YRB) from 2010 to 2019, and the two-way fixed-effects model is employed to illustrate the driving mechanisms of various influencing factors on the RCD in YRB and YREB. The results show that:(1)the RCD levels of YRB and YREB show a fluctuating upward trend during 2010 and 2019, however, both regions have low RCD levels, as seen by the mean RCD indices for YREB and YRB, which are only 0.433 and 0.309, respectively. (2) The RCD level of YREB is higher than that of YEB. In 2019, the "coordinated" provinces in YRB and YREB account for 37.50% and 81.82% of the total number of provinces in the basins, respectively, the "uncoordinated" and "low coordinated" provinces all located in YRB. (3) The RCD of YRB and YREB is significantly improved by REC, URC and RAEC, but not significantly positively by MASCC or EASC, and insufficient development of MASCC is the main contradiction limiting the increase in the RCD level of YRB, while the low level of EASC has become the main obstacle limiting the RCD of YREB. (4)Finally, based on the varying impact degrees and directions of different influencing factors on the RCD in YRB and YREB, the recommendations to promote RCD are proposed.

Identification of runner fatigue stages based on inertial sensors and deep learning.

Introduction: Running is one of the most popular sports in the world, but it also increases the risk of injury. The purpose of this study was to establish a modeling approach for IMU-based subdivided action pattern evaluation and to investigate the classification performance of different deep models for predicting running fatigue. Methods: Nineteen healthy male runners were recruited for this study, and the raw time series data were recorded during the pre-fatigue, mid-fatigue, and post-fatigue states during running to construct a running fatigue dataset based on multiple IMUs. In addition to the IMU time series data, each participant's training level was monitored as an indicator of their level of physical fatigue. Results: The dataset was examined using single-layer LSTM (S_LSTM), CNN, dual-layer LSTM (D_LSTM), single-layer LSTM plus attention model (LSTM + Attention), CNN, and LSTM hybrid model (LSTM + CNN) to classify running fatigue and fatigue levels. Discussion: Based on this dataset, this study proposes a deep learning model with constant length interception of the raw IMU data as input. The use of deep learning models can achieve good classification results for runner fatigue recognition. Both CNN and LSTM can effectively complete the classification of fatigue IMU data, the attention mechanism can effectively improve the processing efficiency of LSTM on the raw IMU data, and the hybrid model of CNN and LSTM is superior to the independent model, which can better extract the features of raw IMU data for fatigue classification. This study will provide some reference for many future action pattern studies based on deep learning.

Experimental periodontitis induced hypoadiponectinemia by IRE1α-mediated endoplasmic reticulum stress in adipocytes.

BACKGROUD: Hypoadiponectinemia is the important cause of insulin resistance. Recent studies have shown that periodontitis is associated with hypoadiponectinemia. The purpose of this study was to investigate the effect of periodontitis-induced endoplasmic reticulum stress (ERS) in visceral adipocytes on hypoadiponectinemia. METHODS: Rat periodontitis models were established by local ligation with silk around the bilateral maxillary second molars. Porphyromonas gingivalis-lipopolysaccharid (P.g-LPS) was also used to stimulate the visceral adipocytes in vitro. The protein expression levels of glucose regulated protein 78 (GRP78), inositol-requiring protein 1α (IRE1α), protein kinase RNA-like ER kinase (PERK), activating transcription factor 6 (ATF6) and adiponectin were detected. IRE1α lentiviruses were transfected into visceral adipocytes in vitro, and an IRE1α inhibitor (KIRA6) was injected in epididymal adipose tissue of rats to detect and verify the effect of ERS on adiponectin expression in visceral adipocytes in vivo. RESULTS: Hypoadiponectinemia was observed in periodontitis rat, and the expression levels of ERS key proteins GRP78 and the phosphorylation levels of IRE1α (p-IRE1α)/IRE1α in visceral adipocytes were increased, while the expression levels of adiponectin protein were decreased. After KIRA6 injection into epididymal adipose tissue of rats with periodontitis, adiponectin levels in visceral adipocytes increased, and serum adiponectin levels recovered to a certain extent. The protein expression levels of GRP78 and p-IRE1α/IRE1α were increased and adiponectin protein expression was decreased in P.g-LPS-induced visceral adipocytes. Overexpression of IRE1α further inhibited adiponectin expression in P.g-LPS-stimulated visceral adipocytes, and conversely, IRE1α inhibition restored adiponectin expression. CONCLUSIONS: Our findings suggest that periodontitis induces ERS in visceral adipocytes leading to hypoadiponectinemia. IRE1α is a key protein regulating adiponectin expression in visceral adipocytes.

Chronic periodontitis induces the proliferation of pancreatic ß-cells to cause hyperinsulinemia in a rat model.

BACKGROUND AND OBJECTIVE: The purpose of this study was to determine if chronic periodontitis (CP) may induce hyperinsulinemia and may have the effect of on pancreatic ß-cell proliferation in a rat model. MATERIALS AND METHODS: Twelve male Sprague-Dawley rats were divided into two groups: the CP group and the control group (Con group). The following contents were evaluated: pathological changes in periodontal soft and hard tissues; serum lipopolysaccharide (LPS) level, serum fasting insulin (FINS) level, fasting blood glucose (FBG) level, and homeostasis model assessment (HOMA) ß (HOMA-ß) index; histopathological examination of islets; immunohistochemistry of insulin and p-Smad2 expression in islets; immunofluorescence of changes in the relative number of ß-cells and the number of Ki67-positive ß-cells. Western blotting was used to analyze p-Smad2/Smad2 levels. Results were analyzed by two independent samples t tests. RESULTS: Increased serum LPS level, FINS level, and HOMA-ß index were observed in the rats of the CP group; FBG level did not change significantly; histological assessments showed an enlarged islet area, increased insulin content, relatively increased ß-cells, increased Ki67-positive ß-cells, and decreased p-Smad2 expression in islets in the rats of the CP group. CONCLUSION: Our study results link CP-induced hyperinsulinemia with changes in islets, such as islet hyperplasia and compensatory ß-cell proliferation, by using a CP rat model.

Effects of rest interval and training intensity on jumping performance: a systematic review and meta-analysis investigating post-activation performance enhancement.

Background: There is no clear agreement regarding the ideal rest interval and training intensity to optimize post-activation performance enhancement (PAPE) after barbell squat (BS). Therefore, the aim of this study was to investigate the effects of rest interval and training intensity on jumping performance due to PAPE. Methods: Searches were performed in PubMed, Web of Science, and EBSCO databases. We included studies that satisfied the following criteria: (1) eligible studies should be randomized controlled trials; (2) eligible studies should investigate the acute effect of barbell squat-induced PAPE on jump performance; (3) eligible studies should use countermovement jump, squat jump, or vertical jump as the outcome measure. Results: From 2518 search records initially identified, 19 studies were eligible for meta-analysis. Our meta-analysis results showed that BS had no significant effect on improving jumping performance due to PAPE (Cohen's d = 0.09, p = 0.08). Subgroup analysis showed that rest interval between 0-1 min was detrimental to jumping performance (Cohen's d = -0.33, p < 0.01), while rest intervals between 4-7 min (Cohen's d = 0.31, p < 0.01) and 8-9 min (Cohen's d = 0.26, p = 0.02) provided favorable jumping performance outcomes. In addition, low-intensity and moderate-intensity BS had no significant effect on jump performance, while high-intensity BS showed results consistent with rest interval. Conclusion: Our study indicated that both low-intensity and moderate-intensity BS failed to induce PAPE, and it is recommended that future studies use high-intensity BS to induce PAPE. A rest interval of 4-9 min had a beneficial impact on jump height, and an interval range of 4-7 min seems to be the best rest interval between conditioning activity and jumping performance.

Development of actein derivatives as potent anti-triple negative breast cancer agents.

Actein is a natural triterpenoid glycoside, isolated from the rhizomes of Cimicifuga foetida, which have been demonstrated to be potential in the treatment of breast cancer previously. Herein, we described the design and synthesis of a series of actein derivatives as anti-triple negative breast cancer (TNBC) inhibitors. Of which, the most promising derivative 27 exhibited significant inhibitory activity against human TNBC cell lines HCC1806 and MDA-MB-231, with IC50 values of 2.78 and 9.11 µM, respectively. Structure-activity relationships of actein derivatives were also discussed. Moreover, preliminary mechanism investigation revealed that 27 significantly inhibited cancer cell proliferation via cell cycle arrest at S phase. In addition, western blot analysis showed that the activation of MAPK signaling pathway might contribute to derivative 27 induced cell death. Overall, these results indicate that 27 has the potential to be developed as a lead compound and compounds with the actein scaffold are a promising novel class of inhibitors to treat TNBC.

Effect of continuous aerobic exercise on endothelial function: A systematic review and meta-analysis of randomized controlled trials.

Background: Current research suggests that continuous aerobic exercise can be effective in improving vascular endothelial function, while the effect between different intensities and durations of exercise is unclear. The aim of this study was to explore the effect of different durations and intensities of aerobic exercise on vascular endothelial function in different populations. Methods: Searches were performed in PubMed, Web of Science, and EBSCO databases. We included studies that satisfied the following criteria: 1) randomized controlled trials (RCTs); 2) including both an intervention and control group; 3) using flow-mediated dilation (FMD) as the outcome measure; and 4) testing FMD on the brachial artery. Results: From 3,368 search records initially identified, 41 studies were eligible for meta-analysis. There was a significant effect of continuous aerobic exercise on improving flow-mediated dilation (FMD) [weighted mean difference (WMD), 2.55, (95% CI, 1.93-3.16), p < 0.001]. Specifically, moderate-intensity [2.92 (2.02-3.825), p < 0.001] and vigorous-intensity exercise [2.58 (1.64-3.53), p < 0.001] significantly increased FMD. In addition, a longer duration [<12 weeks, 2.25 (1.54-2.95), p < 0.001; ≥12 weeks, 2.74 (1.95-3.54), p < 0.001], an older age [age <45, 2.09 (0.78-3.40), p = 0.002; 45 ≤ age <60, 2.25 (1.49-3.01), p < 0.001; age ≥60, 2.62 (1.31-3.94), p < 0.001], a larger basal body mass index (BMI) [20 < BMI < 25, 1.43 (0.98-1.88), p < 0.001; 25 ≤ BMI < 30, 2.49 (1.07-3.90), p < 0.001; BMI ≥ 30, 3.05 (1.69-4.42), p < 0.001], and a worse basal FMD [FMD < 4, 2.71 (0.92-4.49), p = 0.003; 4 ≤ FMD < 7, 2.63 (2.03-3.23), p < 0.001] were associated with larger improvements in FMD. Conclusion: Continuous aerobic exercise, especially moderate-intensity and vigorous-intensity aerobic exercise, contributed to improving FMD. The effect of continuous aerobic exercise on improving FMD was associated with duration and participant's characteristics. Specifically, a longer duration, an older age, a larger basal BMI, and a worse basal FMD contributed to more significant improvements in FMD. Systematic Review Registration: [https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=341442], identifier [CRD42022341442].

Design, synthesis, and anti-triple negative breast cancer activity of novel Toosendanin derivatives.

Toosendanin (TSN) is a natural anti-cancer compound that is isolated from the traditional Chinese herbal Melia toosendan Sieb et Zucc. However, the research effect of TSN in the treatment of Triple negative breast cancer (TNBC) is still far from ideal. In this work, we investigated TSN and its derivatives in terms of their actions against MDA-MB-231 and HCC1806 TNBC cell lines. The results indicated that TSN and its derivative 11 showed excellent antitumor activity. Preliminary mechanistic studies showed that both compounds TSN and 11 induced S-phase arrest and G2/M phase cell number decrease in HCC1806 cells. Also, TSN and 11 significantly reduced the protein level of the well-known cancer suppressor gene p53, reduced the phosphorylation of AKT and ERK, and also induced the accumulation of phosphorylated p38 and p21.

The Effect of Aerobic Exercise on Cognitive Function in People with Alzheimer's Disease: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

A growing body of research has examined the effect of aerobic exercise on cognitive function in people with Alzheimer's Disease (AD), but the findings of the available studies were conflicting. The aim of this study was to explore the effect of aerobic exercise on cognitive function in AD patients. Searches were performed in PubMed, Web of Science, and EBSCO databases from the inception of indexing until 12 November 2021. Cochrane risk assessment tool was used to evaluate the methodological quality of the included literature. From 1942 search records initially identified, 15 randomized controlled trials (RCTs) were considered eligible for systematic review and meta-analysis. Included studies involved 503 participants in 16 exercise groups (mean age: 69.2-84 years) and 406 participants (mean age: 68.9-84 years) in 15 control groups. There was a significant effect of aerobic exercise on increasing mini-mental state examination (MMSE) score in AD patients [weighted mean difference (WMD), 1.50 (95% CI, 0.55 to 2.45), p = 0.002]. Subgroup analyses showed that interventions conducted 30 min per session [WMD, 2.52 (95% CI, 0.84 to 4.20), p = 0.003], less than 150 min per week [WMD, 2.10 (95% CI, 0.84 to 3.37), p = 0.001], and up to three times per week [WMD, 1.68 (95% CI, 0.46 to 2.89), p = 0.007] increased MMSE score significantly. In addition, a worse basal cognitive status was associated with greater improvement in MMSE score. Our analysis indicated that aerobic exercise, especially conducted 30 min per session, less than 150 min per week, and up to three times per week, contributed to improving cognitive function in AD patients. Additionally, a worse basal cognitive status contributed to more significant improvements in cognitive function.

Growth hormone in fertility and infertility: Mechanisms of action and clinical applications.

Secreted by the anterior pituitary gland, growth hormone (GH) is a peptide that plays a critical role in regulating cell growth, development, and metabolism in multiple targeted tissues. Studies have shown that GH and its functional receptor are also expressed in the female reproductive system, including the ovaries and uterus. The experimental data suggest putative roles for GH and insulin-like growth factor 1 (IGF-1, induced by GH activity) signaling in the direct control of multiple reproductive functions, including activation of primordial follicles, folliculogenesis, ovarian steroidogenesis, oocyte maturation, and embryo implantation. In addition, GH enhances granulosa cell responsiveness to gonadotropin by upregulating the expression of gonadotropin receptors (follicle-stimulating hormone receptor and luteinizing hormone receptor), indicating crosstalk between this ovarian regulator and the endocrine signaling system. Notably, natural gene mutation of GH and the age-related decline in GH levels may have a detrimental effect on female reproductive function, leading to several reproductive pathologies, such as diminished ovarian reserve, poor ovarian response during assisted reproductive technology (ART), and implantation failure. Association studies using clinical samples showed that mature GH peptide is present in human follicular fluid, and the concentration of GH in this fluid is positively correlated with oocyte quality and the subsequent embryo morphology and cleavage rate. Furthermore, the results obtained from animal experiments and human samples indicate that supplementation with GH in the in vitro culture system increases steroid hormone production, prevents cell apoptosis, and enhances oocyte maturation and embryo quality. The uterine endometrium is another GH target site, as GH promotes endometrial receptivity and pregnancy by facilitating the implantation process, and the targeted depletion of GH receptors in mice results in fewer uterine implantation sites. Although still controversial, the administration of GH during ovarian stimulation alleviates age-related decreases in ART efficiency, including the number of oocytes retrieved, fertilization rate, embryo quality, implantation rate, pregnancy rate, and live birth rate, especially in patients with poor ovarian response and recurrent implantation failure.

Antidiabetic drug metformin suppresses tumorigenesis through inhibition of mevalonate pathway enzyme HMGCS1.

Metformin, an antidiabetic drug, shows some potent antitumor effects. However, the molecular mechanism of metformin in tumor suppression has not been clarified. Here, we provided evidence using in vitro and in vivo data that metformin inhibited mevalonate pathway by downregulation of 3-hydroxy-3-methylglutaryl-CoA synthase 1 (HMGCS1), a key enzyme in this pathway. Our results further demonstrated that metformin downregulated HMGCS1 expression through inhibition of transcription factor nuclear factor E2-related factor 2. In addition, we determined that HMGCS1 was highly expressed in human liver and lung cancer tissues and associated with lower survival rates. In summary, our study indicated that metformin suppresses tumorigenesis through inhibition of the nuclear factor E2-related factor 2-HMGCS1 axis, which might be a potential target in cancer prevention and treatment.

Electrochemical Detection of Alpha-Fetoprotein Based on Black Phosphorus Nanosheets Modification with Iron Ions.

Black phosphorus nanosheets (BPNSs) were synthesized with liquid exfoliation combined with the ultrasonic method and loaded with Fe3+ by simply mixing. The morphology, structure and electrochemical properties of the synthesized Fe3+/BPNSs were characterized by transmission electron microscopy (TEM), atomic force microscopy (AFM), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS) and cyclic voltammetry (CV), etc. The load of Fe3+ can improve the electrochemical performance of BPNSs and enhance the sensitivity of the detection. Additionally, Fe3+/BPNSs display good biocompatibility. In this study, immunosensors based on Fe3+/BPNSs were constructed to detect alpha-fetoprotein (AFP). The detection is due to the specific binding between the AFP antigen and antibody on the surface of the immunosensors, which can reduce the current response of Fe3+/BPNSs. The immunosensors have a good linear relationship in the range of 0.005 ng·mL-1 to 50 ng·mL-1, and the detection limit is 1.2 pg·mL-1. The results show that surface modification with metal ions is a simple and effective way to improve the electrochemical properties of BPNSs, which will broaden the prospects for the future application of BPNSs in the electrochemical field.

Mechanism of Porphyromonas gingivalis-lipopolysaccharide in regulating the insulin signaling pathway in adipocytes via X-box binding protein 1.

OBJECTIVES: This study aims to investigate the effect of X-box binding protein 1 (XBP1), a key signal molecule of ERS, on the insulin signaling pathway in adipocytes stimulated by Porphyromonas gingivalis (P. gingivalis)-lipopolysaccharide (LPS), a pathogenic bacterium of periodontitis. METHODS: Primary cultured rat adipocytes were stimulated by P. gingivalis-LPS (100 ng·mL-1) for 4, 8, 12, and 24 h. The protein expression levels of insulin receptor substrate-1 (IRS-1), phosphoinositide dependent protein kinase (p-PDK-1), and protein kinase B (p-AKT-1) in the insulin signaling pathway were detected by Western blot analysis. pLVX-NC1, pLVX-XBP1, pLVX-NC2, and pLVX-XBP1-RNAi were transfected into adipocytes, respectively. The transfected rat adipocytes were stimulated by P. gingivalis-LPS, and the protein expression of the insulin signaling pathway was detected by Western blot. RESULTS: The Western Blot showed decreased protein expression of the insulin signaling pathway in rat adipocytes stimulated with P. gingivalis-LPS compared with the control, and the difference was statistically significant (P<0.05). The protein expression levels of IRS-1, p-PDK-1, and p-AKT in the rat adipocytes of pLVX-XBP1 were significantly higher than those in pLVX-NC1 at 8 and 12 h after P. gingivalis-LPS stimulation (P<0.05). The protein expression levels of IRS-1, p-PDK-1, and p-AKT in the rat adipocytes of pLVX-XBP1-RNAi were significantly lower than those in pLVX-NC2 at 4, 8, 12, and 24 h after P. gingivalis-LPS stimulation (P<0.05). CONCLUSIONS: P. gingivalis-LPS regulates the insulin signaling pathway in adipocytes th-rough XBP1.

The Role and Mechanism of Oxidative Stress and Nuclear Receptors in the Development of NAFLD.

The overproduction of reactive oxygen species (ROS) and consequent oxidative stress contribute to the pathogenesis of acute and chronic liver diseases. It is now acknowledged that nonalcoholic fatty liver disease (NAFLD) is characterized as a redox-centered disease due to the role of ROS in hepatic metabolism. However, the underlying mechanisms accounting for these alternations are not completely understood. Several nuclear receptors (NRs) are dysregulated in NAFLD, and have a direct influence on the expression of a set of genes relating to the progress of hepatic lipid homeostasis and ROS generation. Meanwhile, the NRs act as redox sensors in response to metabolic stress. Therefore, targeting NRs may represent a promising strategy for improving oxidation damage and treating NAFLD. This review summarizes the link between impaired lipid metabolism and oxidative stress and highlights some NRs involved in regulating oxidant/antioxidant turnover in the context of NAFLD, shedding light on potential therapies based on NR-mediated modulation of ROS generation and lipid accumulation.

The serine proteases dipeptidyl-peptidase 4 and urokinase are key molecules in human and mouse scar formation.

Despite recent advances in understanding skin scarring, mechanisms triggering hypertrophic scar formation are still poorly understood. In the present study, we investigate mature human hypertrophic scars and developing scars in mice at single cell resolution. Compared to normal skin, we find significant differences in gene expression in most cell types present in scar tissue. Fibroblasts show the most prominent alterations in gene expression, displaying a distinct fibrotic signature. By comparing genes upregulated in murine fibroblasts during scar development with genes highly expressed in mature human hypertrophic scars, we identify a group of serine proteases, tentatively involved in scar formation. Two of them, dipeptidyl-peptidase 4 (DPP4) and urokinase (PLAU), are further analyzed in functional assays, revealing a role in TGFß1-mediated myofibroblast differentiation and over-production of components of the extracellular matrix in vitro. Topical treatment with inhibitors of DPP4 and PLAU during scar formation in vivo shows anti-fibrotic activity and improvement of scar quality, most prominently after application of the PLAU inhibitor BC-11. In this study, we delineate the genetic landscape of hypertrophic scars and present insights into mechanisms involved in hypertrophic scar formation. Our data suggest the use of serine protease inhibitors for the treatment of skin fibrosis.

A retrospective study of endodontic microsurgery about 302 patients.

OBJECTIVES: To investigate the outcome of endodontic microsurgery and analyze the potential prognostic factors, and to evaluate the value of surgical classification by Kim and Kratchman. METHODS: Collecting clinical examination and radiographical examination of endodontic microsurgery cases (which were followed up at least 1 year), which were classified according to Kim and Kratchman, and we analyzed the outcome of endodontic microsurgery and its potential prognostic factors. RESULTS: 302 patients (400 teeth) who received endodontic microsurgery were included. The one year success rate of endodontic microsurgery was 94.25%. Different classification had significant influences on the outcome of endodontic microsurgery (P<0.05), and the success rate of class B and C were better than those of class D, E, and F. The position of teeth had significant influences on the outcome of endodontic microsurgery (P<0.05). The success rate of maxillary teeth was higher than that of mandibular teeth. The success rate of anterior teeth was higher than that of posterior teeth (P<0.05). The quality of root end filling and first or second surgery had no effect on the outcome (P>0.05). CONCLUSIONS: Endodontic microsurgery is an effective treatment method for saving affected teeth, and it can get a good result. Tooth position and classification are the potential prognostic factors. The surgical classification of Kim and Kratchman can help to predict the outcome of endodontic microsurgery.

Twin reversed arterial perfusion syndrome in a monochorionic monoamniotic twin pregnancy.

OBJECTIVE: Twin-Reversed Arterial Perfusion (TRAP) sequence is a rare complication of monochorionic multiple gestation. Conservative management should be considered if there is no poor prognostic factor. CASE REPORT: This is a 35 year-old female with twin pregnancy with acardiac monster. Under the request of the patient, there was no intervention during the whole pregnancy. We keep regular and close sonography weekly follow up. There was no maternal complication and there was also no heart failure sign or polyhydramnios of the donor twin. Minimal blood flow was noted at the anastomotic vessels under the sonography at late gestational age. Due to breech presentation, cesarean section was performed at gestational age 37 + 1/7 weeks. She delivers a healthy baby smoothly. CONCLUSION: Antenatal sonography is an important tool to evaluate the fetus status. Under special condition, term pregnancy is still possible without any treatment. CASE REPORT: Twin reversed arterial perfusion syndrome in a monochorionic monoamniotic twin pregnancy.

N6 -Methyladenosine Reader Protein YT521-B Homology Domain-Containing 2 Suppresses Liver Steatosis by Regulation of mRNA Stability of Lipogenic Genes.

BACKGROUND AND AIMS: Nonalcoholic fatty liver disease (NAFLD) is characterized by accumulation of excessive triglycerides (TGs) in hepatocytes. Obesity is a major risk factor for developing fatty liver, although the intracellular molecular basis remains largely unclear. N6 -methyladenosine (m6 A) RNA methylation is the most common internal modification in eukaryotic mRNA. APPROACH AND RESULTS: In the present study, by m6 A sequencing and RNA sequencing, we found that both m6 A enrichment and mRNA expression of lipogenic genes were significantly increased in leptin-receptor-deficient db/db mice. Importantly, our results showed that YT521-B homology domain-containing 2 (Ythdc2), an m6 A reader, was markedly down-regulated in livers of obese mice and NAFLD patients. Suppression of Ythdc2 in livers of lean mice led to TG accumulation, whereas ectopic overexpression of Ythdc2 in livers of obese mice improved liver steatosis and insulin resistance. Mechanistically, we found that Ythdc2 could bind to mRNA of lipogenic genes, including sterol regulatory element-binding protein 1c, fatty acid synthase, stearoyl-CoA desaturase 1, and acetyl-CoA carboxylase 1, to decrease their mRNA stability and inhibit gene expression. CONCLUSIONS: Our findings describe an important role of the m6 A reader, Ythdc2, for regulation of hepatic lipogenesis and TG homeostasis, which might provide a potential target for treating obesity-related NAFLD.

Screening and identification of epithelial-to-mesenchymal transition-related circRNA and miRNA in prostate cancer.

Epithelial-to-mesenchymal transition (EMT) plays a vital role in the progression and metastasis of prostate cancer. However, the molecular mechanisms underlying prostate cancer metastasis are not fully demonstrated. In this study, EMT was induced by interferon-γ (IFN-γ) in PC-3M IE8 cells. High-throughput sequencing was used to screen the differentially expressed circular RNAs (circRNAs) and miRNAs in the cells with or without IFN-γ treatment. EMT-related circRNAs and miRNAs were further identified by quantitative real-time PCR (qPCR). In addition, the relationships among circRNAs, miRNAs, and mRNA were predicted. After cells were treated with IFN-γ, western blot analysis was conducted to detect the expression levels of EMT markers. E-cadherin expression levels were found to be downregulated, and Twist expression levels were found to be upregulated. Our results also found that IFN-γ promoted PC-3M IE8 cell migration and invasion, indicating that IFN-γ could induce EMT in PC-3M IE8 cells. Furthermore, high-throughput sequencing results revealed 827 upregulated and 1279 downregulated circRNAs and 39 upregulated and 2076 downregulated miRNAs in the IFN-γ group compared with the control group. KEGG analysis showed that both differentially expressed circRNAs and differentially expressed miRNAs were enriched in the MAPK signaling pathway related to EMT. Furthermore, the qPCR results revealed that the expression of hsa_circ_0001085, hsa_circ_0004916, hsa_circ_0001165, hsa-miR-196b-5p, and hsa-miR-187-3p in the IFN-γ group was consistent with the sequencing results. hsa_circ_0001165 and hsa_circ_0001085 were used to construct the network of circRNA-miRNA-mRNA. It was found that hsa_circ_0001165 may regulate TNF expression through hsa-miR-187-3p to induce EMT in prostate cancer cells. In addition, hsa_circ_0001085 may indirectly regulate the PI3K-Akt signaling and TGF-ß signaling pathways through hsa-miR-196b-5p and the MAPK signaling pathway through has-miR-451a, which played a regulatory role in prostate cancer cells in the EMT induction model. The results obtained in this study lay the foundation for future study.