A golgi targeting viscosity rotor for cancer diagnosis in living cells and tissues.

Unveiling the intricate relationship between cancer and Golgi viscosity remains an arduous endeavor, primarily due to the lack of Golgi-specific fluorescent probes tailored for viscosity measurement. Considering this formidable obstacle, we have triumphed over the challenge by devising a bespoke Golgi-specific viscosity probe, aptly named GOL-V. This ingenious innovation comprises the viscosity rotor BODIPY intricately tethered to the Golgi-targeting moiety benzsulfamide. GOL-V exhibits remarkable sensitivity to fluctuations in viscosity, the fluorescence intensity of GOL-V increased 114-fold when the viscosity value was increased from 2.63 to 937.28 cP. Owing to its remarkable capacity to suppress the TICT state under conditions of heightened viscosity. Moreover, its efficacy in sensitively monitoring Golgi viscosity alterations within living cells is also very significant. Astonishingly, our endeavors have culminated in not only the visualization of Golgi viscosity at the cellular and tissue levels but also in the clinical tissue samples procured from cancer patients. Harnessing the prowess of GOL-V, we have successfully demonstrated that Golgi viscosity could serve as a discerning marker for detecting the presence of cancer. The convergence of these exceptional attributes firmly establishes GOL-V as an immensely potent instrument, holding immense potential in the realm of cancer diagnosis.

LncRNA GAS5 restrains ISO-induced cardiac fibrosis by modulating mir-217 regulation of SIRT1.

Considering the effect of SIRT1 on improving myocardial fibrosis and GAS5 inhibiting occurrence and development of myocardial fibrosis at the cellular level, the aim of the present study was to investigate whether LncRNA GAS5 could attenuate cardiac fibrosis through regulating mir-217/SIRT1, and whether the NLRP3 inflammasome activation was involved in this process. Isoprenaline (ISO) was given subcutaneously to the male C57BL/6 mice to induce myocardial fibrosis and the AAV9 vectors were randomly injected into the left ventricle of each mouse to overexpress GAS5. Primary myocardial fibroblasts (MCFs) derived from neonatal C57BL/6 mice and TGF-ß1 were used to induce fibrosis. And the GAS5 overexpressed MCFs were treated with mir-217 mimics and mir-217 inhibitor respectively. Then the assays of expression levels of NLRP3, Caspase-1, IL-1ß and SIRT1 were conducted. The findings indicated that the overexpression of GAS5 reduced the expression levels of collagen, NLRP3, Capase-1, IL-1ß and SIRT1 in ISO treated mice and TGF-ß1 treated MCFs. However, this effect was significantly weakened after mir-217 overexpression, but was further enhanced after knockdown of mir-217. mir-217 down-regulates the expression of SIRT1, leading to increased activation of the NLRP3 inflammasome and subsequent pyroptosis. LncRNA GAS5 alleviates cardiac fibrosis induced via regulating mir-217/SIRT1 pathway.

[Retracted] Overexpression of indoleamine 2, 3­dioxygenase contributes to the repair of human airway epithelial cells inhibited by dexamethasone via affecting the MAPK/ERK signaling pathway.

[This retracts the article DOI: 10.3892/etm.2018.6163.].

Hydrogen sulfide ameliorates lipopolysaccharide-induced anxiety-like behavior by inhibiting checkpoint kinase 1 activation in the hippocampus of mice.

Hydrogen sulfide (H2S), an endogenous gasotransmitter, exhibits the anxiolytic roles through its anti-inflammatory effects, although its underlying mechanisms remain largely elusive. Emerging evidence has documented that cell cycle checkpoint kinase 1 (Chk1)-regulated DNA damage plays an important role in the neurodegenerative diseases; however, there are few relevant reports on the research of Chk1 in neuropsychiatric diseases. Here, we aimed to investigate the regulatory role of H2S on Chk1 in lipopolysaccharide (LPS)-induced anxiety-like behavior focusing on inflammasome activation in the hippocampus. Cystathionine γ-lyase (CSE, a H2S-producing enzyme) knockout (CSE-/-) mice displayed anxiety-like behavior and activation of inflammasome-mediated inflammatory responses, manifesting by the increase levels of interleukin-1ß (IL-1ß), IL-6, and ionized calcium-binding adaptor molecule-1 (Iba-1, microglia marker) expression in the hippocampus. Importantly, expression of p-Chk1 and γ-H2AX (DNA damage marker) levels were also increased in the hippocampus of CSE-/- mice. LPS treatment decreased the expression of CSE and CBS while increased p-Chk1 and γ-H2AX levels and inflammasome-activated neuroinflammation in the hippocampus of mice. Moreover, p-Chk1 and γ-H2AX protein levels and cellular immunoactivity were significantly increased while CSE and CBS were markedly decreased in cultured BV2 cells followed by LPS treatment. Treatment of mice with GYY4137, a donor of H2S, inhibited LPS-induced increased in p-Chk1 and γ-H2AX levels, mitigated inflammasome activation and inflammatory responses as well as amelioration of anxiety-like behavior. Notably, SB-218078, a selective Chk1 inhibitor treatment attenuated the effect of LPS on inflammasome activation and inflammatory responses and the induction of anxiety-like behavior. Finally, STAT3 knockdown with AAV-STAT3 shRNA alleviated LPS-induced anxiety-like behavior and inhibited inflammasome activation in the hippocampus, and blockade of NLRP3 with MCC950 attenuated neuroinflammation induction and ameliorated LPS-induced anxiety-like behavior. Overall, this study indicates that downregulation of Chk1 activity by H2S activation may be considered as a valid strategy for preventing the progression of LPS-induced anxiety-like behavior.

Association between white blood cell count and adverse pregnancy outcomes: a retrospective cohort study from a tertiary hospital in China.

OBJECTIVES: This study aimed to clarify the relationship between white blood cell (WBC) and adverse pregnancy outcomes. DESIGN: A total of 25 270 pregnant women underwent peripheral blood white blood cell count tests in the first, second and third trimesters. Adverse pregnancy outcomes were gestational hypertension, pre-eclampsia, gestational diabetes mellitus, preterm birth, low birth weight, caesarean delivery, macrosomia and fetal distress. Due to acute infectious disease or other diseases, 1127 were excluded. SETTING: Minhang Hospital, China. PARTICIPANTS: A total of 24 143 pregnant women were included in this study. PRIMARY AND SECONDARY OUTCOME MEASURES: The primary outcome was the adverse pregnancy outcomes. RESULTS: For the 24 143 participants, we calculated adjusted ORs for adverse pregnancy outcomes associated with an increased WBC count. For gestational hypertension, the ORs were 1.18 (95% CI, 1.05 to 1.24) in the first trimester and 1.10 (1.06 to 1.13) in the second trimester; for pre-eclampsia, ORs were 1.14 (95% CI, 1.47 to 1.64) in the first trimester and 1.10 (1.05 to 1.16) in the second trimester; for gestational diabetes mellitus, ORs were 1.06 (95% CI, 1.00 to 1.13) in the first trimester and 1.10 (1.04 to 1.16) in the second trimester; for preterm birth, ORs were 1.12 (95% CI, 1.06 to 1.18) in the first trimester, 1.10 (1.06 to 1.13) in the second trimester and 1.12 (1.09 to 1.15) in the third trimester; for low birth weight, ORs were 1.09 (95% CI, 1.02 to 1.17) in the first trimester, 1.03 (0.99 to 1.08) in the second trimester and 1.12 (1.08 to 1.16) in the third trimester. Significant associations were not observed obviously for caesarean delivery, macrosomia and fetal distress. CONCLUSIONS: Our results indicate strong, continuous associations of maternal WBC count with increased risks of adverse pregnancy outcomes.

The diagnostic value of ultrasound on different-sized thyroid nodules based on ACR TI-RADS.

OBJECTIVES: The thyroid nodule is one of the most common endocrine system diseases. Risk classification models based on ultrasonic features have been created by multiple professional societies, including the American College of Radiology (ACR), which published the Thyroid Imaging Reporting and Data System (TI-RADS) in 2017. The effect of the size in the diagnostic value of ultrasound remains not well defined. The purposes of our study aims to explore diagnostic value of the ACR TI-RADS on different-sized thyroid nodules. METHODS: A total of 1183 thyroid nodules were selected from 952 patients with thyroid nodules confirmed by surgical pathology from January 2021 to October 2022. Based on the maximum diameters of the nodules, they were stratified into groups A ( ≤ 10 mm), B ( > 10 mm, < 20 mm) and C ( ≥ 20 mm). The ultrasonic features of the thyroid nodules in each group were evaluated and scored based on ACR TI-RADS, and the receiver operating characteristic curve (ROC) was plotted to determine the optimal cut-off value for the ACR TI-RADS scores and categories in each group. Finally, the diagnostic efficacy of ACR TI-RADS on different-sized thyroid nodules was analyzed. RESULTS: Among the 1183 thyroid nodules, 340 were benign, 10 were low-risk and 833 were malignant. For the convenience of statistical analysis, low-risk thyroid nodules were classified as malignant in this study. The ACR TI-RADS scores and categorical levels of malignant thyroid nodules in each group were higher than those of benign ones (p < 0.05). The areas under the ROCs (AUCs) plotted based on scores were 0.741, 0.907, and 0.904 respectively in the three groups, and the corresponding optimal cut-off values were > 6 points, > 5 points and > 4 points respectively. While the AUCs of the ACR TI-RADS categories were 0.668, 0.855, and 0.887 respectively in each group, with the optimal cut-off values were all > TR4. Besides, for thyroid nodules of larger sizes, ACR TI-RADS exhibited weaker sensitivity with lower positive prediction value (PPV), but the specificity and negative prediction value (NPV) were both higher, presenting with statistically significant differences (p < 0.05). CONCLUSION: For thyroid nodules of different sizes, the diagnostic efficacy of ACR TI-RADS varies as well. The system shows better diagnostic efficacy on thyroid nodules of > 10 mm than on those ≤ 10 mm. Considering the favorable prognosis of thyroid microcarcinoma and the low diagnostic efficacy of ACR TI-RADS on it, the scoring and classification of thyroid micro-nodules can be left out in appropriate cases, so as to avoid the over-diagnosis and over-treatment of thyroid microcarcinoma to a certain extent.

Comparing Diagnostic Efficacy of C-TIRADS Positive Features on Different Sizes of Thyroid Nodules.

Purpose: To explore the diagnostic value of positive features in the Chinese Thyroid Imaging Reporting and Data System (C-TIRADS) for thyroid nodules of different sizes. Patients and Methods: A total of 1864 patients with 2347 thyroid nodules were selected from January 2021 to December 2022 and assessed according to C-TIRADS. According to the maximum diameter, nodules were divided into the A1 group (≤10 mm), A2 group (>10 mm,<20 mm), and A3 group (≥20 mm). With surgical pathology as the golden standard, the receiver operating characteristic curves (ROC) were constructed, and each group's area under the curve (AUC) was calculated. The diagnostic value of positive features in C-TIRADS for different sizes of thyroid nodules was analyzed. Results: In all groups, malignant thyroid nodules had a higher incidence of positive features than benign nodules (P < 0.05). In A1 group, the diagnostic efficiency of C-TIRADS positive features for thyroid nodules was vertical orientation> ill-defined/irregular margin or extrathyroidal extension> solid composition> markedly hypoechoic> microcalcifications. The AUCs were 0.718, 0.675, 0.609, 0.558, and 0.581, respectively. In A2 group, the diagnostic efficacy of each positive features for thyroid nodules was ill-defined/irregular margins or extra-thyroid invasion> solid composition> microcalcifications> markedly hypoechoic> vertical orientation. The AUCs were 0.854, 0.730, 0.719, 0.670, and 0.609, respectively. In A3 group, the diagnostic efficacy of each positive features for thyroid nodules was ill-defined/irregular margin or extrathyroidal extension> microcalcifications> solid composition> vertical orientation> markedly hypoechoic. The AUCs were 0.847, 0.778, 0.767, 0.584, and 0.560, respectively. Conclusion: C-TIRADS positive features exhibited different diagnostic efficacy for thyroid nodules of various sizes, especially for thyroid nodules ≤10 mm, for which all positive features had low diagnostic efficacy.

VGLL4 promotes vascular endothelium specification via TEAD1 in the vascular organoids and human pluripotent stem cells-derived endothelium model.

BACKGROUND: We have shown that Hippo-YAP signaling pathway plays an important role in endothelial cell differentiation. Vestigial-like family member 4 (VGLL4) has been identified as a YAP inhibitor. However, the exact function of VGLL4 in vascular endothelial cell development remains unclear. In this study, we investigated the role of VGLL4, in human endothelial lineage specification both in 3D vascular organoid and 2D endothelial cell differentiation. METHODS AND RESULTS: In this study, we found that VGLL4 was increased during 3D vascular organoids generation and directed differentiation of human embryonic stem cells H1 towards the endothelial lineage. Using inducible ectopic expression of VGLL4 based on the piggyBac system, we proved that overexpression of VGLL4 in H1 promoted vascular organoids generation and endothelial cells differentiation. In contrast, VGLL4 knockdown (heterozygous knockout) of H1 exhibited inhibitory effects. Using bioinformatics analysis and protein immunoprecipitation, we further found that VGLL4 binds to TEAD1 and facilitates the expression of endothelial master transcription factors, including FLI1, to promote endothelial lineage specification. Moreover, TEAD1 overexpression rescued VGLL4 knockdown-mediated negative effects. CONCLUSIONS: In summary, VGLL4 promotes EC lineage specification both in 3D vascular organoid and 2D EC differentiation from pluripotent stem cell, VGLL4 interacts with TEAD1 and facilitates EC key transcription factor, including FLI1, to enhance EC lineage specification.

Rare Vulvar Angiomyofibroblastoma: Case Series and Literature Review.

Background: Angiomyofibroblastoma (AMFB) is an uncommon disease with few literature reports, leading to the poor understanding of its diagnosis, treatment, and postoperative follow-up plans among gynecologists. Objective: To study the clinical and pathological features of vulvar AMFB and discuss its treatment and prognosis. Case Summary: The 3 cases were characterized by a gradually increasing painless mass in the vulva. Preoperative diagnosis was difficult and mainly depended on ultrasonic examination. Immunohistochemistry confirmed clear boundaries of AMFB. This condition could be completely cured by surgery, and the prognosis was good. Conclusion: The vulvar AMFB is a rare tumor that is frequently misdiagnosed before surgery. Ultrasound is preferred in auxiliary diagnosis, and surgery remains the best treatment, and long-term follow-up is necessary to avoid recurrence or other complications.

VGLL4-TEAD1 promotes vascular smooth muscle cell differentiation from human pluripotent stem cells via TET2.

The Hippo signaling pathway plays a critical role in cardiovascular development and stem cell differentiation. Using microarray profiling, we found that the Hippo pathway components vestigial-like family member 4 (VGLL4) and TEA domain transcription factor 1 (TEAD1) were upregulated during vascular smooth muscle cell (VSMC) differentiation from H1 ESCs (H1 embryonic stem cells). To further explore the role and molecular mechanisms of VGLL4 in regulating VSMC differentiation, we generated a VGLL4-knockdown H1 ESC line (heterozygous knockout) using the CRISPR/Cas9 system and found that VGLL4 knockdown inhibited VSMC specification. In contrast, overexpression of VGLL4 using the PiggyBac transposon system facilitated VSMC differentiation. We confirmed that this effect was mediated via TEAD1 and VGLL4 interaction. In addition, bioinformatics analysis revealed that Ten-eleven-translocation 2 (TET2), a DNA dioxygenase, is a target of TEAD1, and a luciferase assay further verified that TET2 is the target of the VGLL4-TEAD1 complex. Indeed, TET2 overexpression promoted VSMC marker gene expression and countered the VGLL4 knockdown-mediated inhibitory effects on VSMC differentiation. In summary, we revealed a novel role of VGLL4 in promoting VSMC differentiation from hESCs and identified TET2 as a new target of the VGLL4-TEAD1 complex, which may demethylate VSMC marker genes and facilitate VSMC differentiation. This study provides new insights into the VGLL4-TEAD1-TET2 axis in VSMC differentiation and vascular development.

Nonalcoholic Fatty Liver Disease and Adverse Pregnancy Outcomes in Women With Normal Prepregnant Weight.

CONTEXT: Existing studies focusing on the effects of nonalcoholic fatty liver disease (NAFLD) combined with normal prepregnant weight on pregnancy outcomes are limited. OBJECTIVE: This study aimed to explore the relationship between maternal NAFLD and adverse pregnancy outcomes in different body mass index (BMI) groups. METHODS: Using an antenatal care and delivery database, we retrospectively analyzed women who delivered in Minhang Hospital affiliated to Fudan University, Shanghai, China from January 1, 2013, to June 30, 2020. NAFLD was confirmed by ultrasound in early pregnancy. A logistic regression model with adjustment for confounders was used to examine potential associations between NAFLD and pregnancy outcomes. RESULTS: A total of 14 708 pregnant women (mean prepregnant BMI 21.0 [SD, 2.8] kg/m2) were included in our final study, of whom 554 (3.8%) had NAFLD. After fully adjusting for potential confounders, NAFLD significantly increased the risk of gestational diabetes mellitus (adjusted odds ratio 2.477; 95% CI, 1.885-3.254), gestational hypertension (3.054; 2.191-4.257), preeclampsia/eclampsia (3.994; 2.591-6.005), cesarean section (1.569; 1.315-1.872), preterm births (1.831; 1.229-2.727), and macrosomia (1.691; 1.300-2.198). It is notable that 83.9% (12 338) of women were of normal weight at the start of pregnancy (prepregnant 18.5 ≤ BMI < 24 kg/m2), and they still had higher odds of adverse pregnancy outcomes. CONCLUSION: Women with NAFLD and a normal weight have a higher risk for adverse pregnancy outcomes. Pregnant women with NAFLD, regardless of obesity status, should be offered a more qualified surveillance to optimize pregnancy outcomes.

Role of Ero1α in cognitive impairment induced by chronic hypoxia.

Recent reports suggested the endoplasmic reticulum stress (ERS)-associated pathway is involved with cognitive impairment in hypoxia condition. ERO1-like protein alpha (Ero1α), an endoplasmic reticulum membrane-bound N-glycoprotein, has been reported to promote oxidative protein folding. However, no studies have reported whether the Ero1α is trapped in hypoxia-induced neuronal loss through the ERS-associated pathways. In our study, this effect of Ero1α was investigated using C57BL/6J mice, the HT22 cells and primary rat neurons. C57BL/6J mice were modeled in a hypoxic chamber for 4 weeks. Behavioral tests were then carried out to test cognitive functions, including the Morris water maze and fear conditioning test. Proteomics showed that Ero1α distinctly upregulated compared with normoxia group and verified using western blotting. Flow cytometry and immunofluorescence were used to analyze the neuroprotective effect of inhibitor EN460 of Ero1α in the HT22 cells. In C57BL/6J mice, hypoxia significantly caused cognitive decline. Brain slice staining results were also used to confirm this effect. Western blot analysis demonstrated that Ero1α, ERS-associated proteins and apoptosis-associated proteins significantly increased in the hypoxia treated groups, further proliferation-related marker protein decreased. EN460, a selective endoplasmic reticulum oxidation 1 (ERO1) inhibitor, counteracted neuronal apoptosis and ameliorated neuronal cell proliferation in the HT22 cells. Taken together, our data indicate that hypoxia induces cognitive impairment, at least in part, by upregulating Ero1α which contributes to neuronal apoptosis through ERS signaling pathway, providing preliminary experimental evidence that the Ero1α is a promising therapeutic target in hypoxia-induced cognitive deficits.

Case Report: Rhabdomyolysis secondary to vildagliptin overdose in a suicidal attempt: A case report and brief literature review.

Rhabdomyolysis is a life-threatening syndrome associated with direct or indirect muscle damage that is rarely reported with dipeptidyl peptidase (DPP)-4 inhibitors. Here we presented a case in which a 58-year-old female suffered from severe swelling and pain in bilateral lower limbs and oliguria after a suicidal vildagliptin overdose. Drug-induced rhabdomyolysis and drug-induced liver injury were diagnosed based on laboratory and radiological findings. The patient was treated with fluid resuscitation, insulin, electrolyte replacement, diuretics, urine alkalizing agents, anticoagulants, antioxidants, and 24-h bedside ECG monitoring and suicide prevention. After 20 days of hospitalization and close monitoring, the patient was discharged without sequelae. Risk factors, diagnostic criteria, disease mechanisms, and outcomes were also discussed. This case illustrated that overdose of oral anti-diabetic medications may result in clinically significant adverse events, such as rhabdomyolysis in this case with a DPP-4 inhibitor. Although the incidence is low, special attention should be paid to intentional or accidental exposure to anti-diabetic medications during suicide attempts, especially in depressed patients with diabetes.

The Apelin-APJ axis alleviates LPS-induced pulmonary fibrosis and endothelial mesenchymal transformation in mice by promoting Angiotensin-Converting Enzyme 2.

Fibrotic alterations resulting from abnormal tissue repair after lung injury are responsible for the high mortality observed after acute respiratory distress syndrome. Therefore, the prevention and treatment of pulmonary fibrosis has been widely concerned. The Apelin-APJ axis plays an important role in the prevention and treatment of respiratory diseases and organ fibrosis. However, its underlying mechanism remains to be further studied. The aim of this study was to investigate whether the anti-pulmonary fibrosis effect of apelin-APJ axis is related to the activation of angiotensin-converting Enzyme 2 (ACE2). Here, we found that exogenous activation of the Apelin-APJ axis alleviates lipopolysaccharide (LPS)-induced pulmonary fibrosis in mice. In vitro studies revealed that Apelin-13 inhibited LPS-induced endothelial mesenchymal transition in lung microvascular endothelial cells, whereas [Ala13]-Apelin-13 (Apelin-APJ axis inhibitor) accelerated LPS-induced endothelial interstitial transformation in lung microvascular endothelial cells. Notably, angiotensin-converting enzyme 2 (ACE2) inhibitor blocks the beneficial effect of the Apelin-APJ axis activation on LPS-induced pulmonary fibrosis. This finding suggests that the Apelin-APJ axis inhibits pulmonary fibrosis by activating ACE2. Simultaneously, accumulating evidence suggests that ubiquitination may contribute to pulmonary fibrosis. Our study found that LPS increased the ubiquitination of ACE2 protein, whereas Apelin-13 inhibited it. In conclusion, exogenous activation of the Apelin-APJ axis improves LPS-induced pulmonary fibrosis in mice and may be a viable therapeutic target for pulmonary fibrosis.

ChK1 activation induces reactive astrogliosis through CIP2A/PP2A/STAT3 pathway in Alzheimer's disease.

Cancerous Inhibitor of PP2A (CIP2A), an endogenous PP2A inhibitor, is upregulated and causes reactive astrogliosis, synaptic degeneration, and cognitive deficits in Alzheimer's disease (AD). However, the mechanism underlying the increased CIP2A expression in AD brains remains unclear. We here demonstrated that the DNA damage-related Checkpoint kinase 1 (ChK1) is activated in AD human brains and 3xTg-AD mice. ChK1-mediated CIP2A overexpression drives inhibition of PP2A and activates STAT3, then leads to reactive astrogliosis and neurodegeneration in vitro. Infection of mouse brain with GFAP-ChK1-AAV induced AD-like cognitive deficits and exacerbated AD pathologies in vivo. In conclusion, we showed that ChK1 activation induces reactive astrogliosis, degeneration of neurons, and exacerbation of AD through the CIP2A-PP2A-STAT3 pathway, and inhibiting ChK1 may be a potential therapeutic approach for AD treatment.

YAP inhibition promotes endothelial cell differentiation from pluripotent stem cell through EC master transcription factor FLI1.

Endothelial cells (ECs) derived from pluripotent stem cells (PSCs) provide great resource for vascular disease modeling and cell-based regeneration therapy. However, the molecular mechanisms of EC differentiation are not completely understood. In this study, we checked transcriptional profile by microarray and found Hippo pathway is changed and the activity of YAP decreased during mesoderm-mediated EC differentiation from human embryonic stem cells (hESCs). Knockdown of YAP in hESCs promoted both mesoderm and EC differentiation indicating by mesodermal- or EC-specific marker gene expression increased both in mRNA and protein level. In contrast, overexpression of YAP inhibited mesoderm and EC differentiation. Microarray data showed that several key transcription factors of EC differentiation, such as FLI1, ERG, SOX17 are upregulated. Interestingly, knockdown YAP enhanced the expression of these master transcription factors. Bioinformation analysis revealed that TEAD, a YAP binds transcription factors, might regulate the expression of EC master TFs, including FLI1. Luciferase assay confirmed that YAP binds to TEAD1, which would inhibit FLI1 expression. Finally, FLI1 overexpression rescued the effects of YAP overexpression-mediated inhibition of EC differentiation. In conclusion, we revealed the inhibitory effects of YAP on EC differentiation from PSCs, and YAP inhibition might promote expression of master TFs FLI1 for EC commitment through interacting with TEAD1, which might provide an idea for EC differentiation and vascular regeneration via manipulating YAP signaling.

[Effects of L-carnitine on autophagy and apoptosis of mouse pulmonary microvascular endothelial cells induced by lipopolysaccharide].

Objective: To investigate the protective effects of L-carnitine (LC) on lipopolysaccharide (LPS) - injured mouse pulmonary microvascular endothelial cells (PMVECs) and its effects on autophagy and apoptosis. Methods: Cultured mouse PMVECs were divided into three groups: ① Control group, ② LPS group (10 µg/ml, 3, 6, 12, 24 h), ③ LPS (10 µg/ml, 24 h)+LC (2.5, 5.0, 10 µg/ml) (LPS+LC) group. PMVECs apoptosis was examined by Annexin V-FITC/PI double labeling method. Autophagosome was detected by immunofluorescence staining. Levels of autophagy-related protein LC3 and apoptosis-related protein caspase-3 were detected by Western blot. PMVECs viability was measured by CCK-8. Results: ① Compared with the control group, LPS treatment inhibited the PMVECs viability significantly, whereas the apoptosis rate and the expression of autophagy protein LC3 II were markedly increased after LPS treatment for 6 h, 12 h and 24 h. ② Compared with LPS group (10 µg/ml, 24 h), the PMVECs viability, levels of autophagy protein LC3 II and caspase-3 protein expression as well as apoptosis rate in LPS+LC group were increased significantly. Conclusion: LC can increase the activity of PMVECs injuried by LPS, promote autophagy and inhibit apoptosis of PMVECs.

Critical role of VGLL4 in the regulation of chronic normobaric hypoxia-induced pulmonary hypertension in mice.

Pulmonary hypertension (PH), a rare but deadly cardiopulmonary disorder, is characterized by extensive remodeling of pulmonary arteries resulting from enhancement of pulmonary artery smooth muscle cell proliferation and suppressed apoptosis; however, the underlying pathophysiological mechanisms remain largely unknown. Recently, epigenetics has gained increasing prominence in the development of PH. We aimed to investigate the role of vestigial-like family member 4 (VGLL4) in chronic normobaric hypoxia (CNH)-induced PH and to address whether it is associated with epigenetic regulation. The rodent model of PH was established by CNH treatment (10% O2 , 23 hours/day). Western blot, quantitative reverse transcription polymerase chain reaction, immunofluorescence, immunoprecipitation, and adeno-associated virus tests were performed to explore the potential mechanisms involved in CNH-induced PH in mice. VGLL4 expression was upregulated and correlated with CNH in PH mouse lung tissues in a time-dependent manner. VGLL4 colocalized with α-smooth muscle actin in cultured pulmonary arterial smooth muscle cells (PASMCs), and VGLL4 immunoactivity was increased in PASMCs following hypoxia exposure in vitro. VGLL4 knockdown attenuated CNH-induced PH and pulmonary artery remodeling by blunting signal transducer and activator of transcription 3 (STAT3) signaling; conversely, VGLL4 overexpression exacerbated the development of PH. CNH enhanced the acetylation of VGLL4 and increased the interaction of ac-H3K9/VGLL4 and ac-H3K9/STAT3 in the lung tissues, and levels of ac-H3K9, p-STAT3/STAT3, and proliferation-associated protein levels were markedly up-regulated, whereas apoptosis-related protein levels were significantly downregulated, in the lung tissues of mice with CNH-induced PH. Notably, abrogation of VGLL4 acetylation reversed CNH-induced PH and pulmonary artery remodeling and suppressed STAT3 signaling. Finally, STAT3 knockdown alleviated CNH-induced PH. In conclusion, VGLL4 acetylation upregulation could contribute to CNH-induced PH and pulmonary artery remodeling via STAT3 signaling, and abrogation of VGLL4 acetylation reversed CNH-induced PH. Pharmacological or genetic deletion of VGLL4 might be a potential target for therapeutic interventions in CNH-induced PH.

Mammography breast density: an effective supplemental modality for the precise grading of ultrasound BI-RADS 4 categories.

BACKGROUND: High breast density is significantly associated with an increased risk of breast diseases. Presently, suspected breast masses assessed as Breast Imaging-Reporting and Data System (BI-RADS) grade 4 provide a wide range of positive predictive values. Moreover, subcategories (4a, 4b, and 4c) are still under consideration as the diagnostic criteria are neither comprehensive nor objective. However, whether mammography breast density (MBD) has any impact on the accurate grading of BI-RADS 4 assessed by ultrasound (US) remains unknown. METHODS: A total of 1,086 women with 1,293 breast masses were included and assessed as BI-RADS 3-5 by US. The subcategories of MBD (from the ACR-a to the ACR-d group) were assessed by mammography according to the criteria of the American College of Radiology (ACR). The clinicopathological characteristics of these patients were reviewed retrospectively. The malignancy rates of breast masses among different subgroups assessed by BI-RADS were re-estimated with MBD. RESULTS: Almost all BI-RADS 3 masses were classified as benign and nearly all BI-RADS 5 masses were identified as malignant. Significant inverse associations between MBD and malignancy rates were detected between the BI-RADS 4a and BI-RADS 4b groups. Moreover, malignancy rates decreased significantly from ACR-a to ACR-d for BI-RADS 4a and 4b breast lesions (P<0.001). However, this trend was not observed in BI-RADS 4c breast lesions. CONCLUSIONS: MBD could serve as a crucial factor for the accurate grading of BI-RADS 4 lesions assessed by US. We strongly recommend the adoption of the MBD as a possible supplemental screening modality for US. Furthermore, it is equally beneficial for accurate risk assessment and screening recommendations based on MBD.