Sprouty genes regulate activated fibroblasts in mammary epithelial development and breast cancer.

Stromal fibroblasts are a major stem cell niche component essential for organ formation and cancer development. Fibroblast heterogeneity, as revealed by recent advances in single-cell techniques, has raised important questions about the origin, differentiation, and function of fibroblast subtypes. In this study, we show in mammary stromal fibroblasts that loss of the receptor tyrosine kinase (RTK) negative feedback regulators encoded by Spry1, Spry2, and Spry4 causes upregulation of signaling in multiple RTK pathways and increased extracellular matrix remodeling, resulting in accelerated epithelial branching. Single-cell transcriptomic analysis demonstrated that increased production of FGF10 due to Sprouty (Spry) loss results from expansion of a functionally distinct subgroup of fibroblasts with the most potent branching-promoting ability. Compared to their three independent lineage precursors, fibroblasts in this subgroup are "activated," as they are located immediately adjacent to the epithelium that is actively undergoing branching and invasion. Spry genes are downregulated, and activated fibroblasts are expanded, in all three of the major human breast cancer subtypes. Together, our data highlight the regulation of a functional subtype of mammary fibroblasts by Spry genes and their essential role in epithelial morphogenesis and cancer development.

Sex-specific differences in the associations between adiposity indices and incident hyperuricemia among middle-aged and older adults: a nationwide longitudinal study.

Objective: Although obesity is a known risk for hyperuricemia (HUA), the associations between adiposity indices and incident HUA and whether sex-specific differences exist is still unknown. We aimed to investigate the associations between adiposity indices and incident HUA in a longitudinal study. Methods: Data from the China Health and Retirement Longitudinal Study (CHARLS) in 2011-2012 and 2015-2016 were used to conduct a cohort study. Participants aged ≥45 years without HUA at baseline were included in this study. Adiposity indices, including body mass index (BMI), waist circumference (WC), waist-to-height ratio body roundness index (BRI), conicity index (CI), lipid accumulation product (LAP) index, waist-to-height ratio (WHtR), visceral adiposity index (VAI), and Chinese visceral adiposity index (CVAI), were calculated. Logistic analysis was used to analyze the association between adiposity indices and incident HUA risk stratified by gender. Receiver operating characteristic curve analysis was performed to evaluate the power of predictions for incident HUA. Results: Of 5,873 participants aged 59.0 ± 8.7 years enrolled in this study, 578 (9.8%) participants developed HUA during the 4-year follow-up period. After adjusting for confounding variables, LAP, VAI, and CVAI showed significant association with incident HUA. BMI, WC, WHtR, BRI, and CI were significantly associated with incident HUA in women but not in men. LAP had the highest area under the curve (AUC) (0.612) followed by CVAI (0.596) in men, while CVAI had the highest AUC (0.707) followed by LAP (0.691) in women. All indices showed better predictive ability in women than in men. Conclusion: Our findings indicated that adiposity indices were effective predictors of incident HUA and showed better predictive power in women than men. In clinical practice, adiposity indices could be used to assess and prevent incident HUA among Chinese middle-aged and older adults.

Chemical reprogramming of melanocytes to skeletal muscle cells.

BACKGROUND: Direct cell-fate conversion by chemical reprogramming is promising for regenerative cell therapies. However, this process requires the reactivation of a set of master transcription factors (TFs) of the target cell type, which has proven challenging using only small molecules. METHODS: We developed a novel small-molecule cocktail permitting robust skin cell to muscle cell conversion. By single cell sequencing analysis, we identified a Pax3 (Paired box 3)-expressing melanocyte population holding a superior myogenic potential outperforming other seven types of skin cells. We further validated the single cell sequencing analysis results using immunofluorescence staining, in situ hybridization and FACS sorting and confirmed the myogenic potential of melanocytes during chemical reprogramming. We used single cell RNA-seq that detect the potential converted cell type, uncovering a unique role of Pax3 in facilitating chemical reprogramming from melanocytes to muscle cells. RESULTS: In this study, we demonstrated that the Pax3-expressing melanocytes to be a skin cell type for skeletal muscle cell fate conversion in chemical reprogramming. By developing a small-molecule cocktail, we showed an efficient melanocyte reprogramming to skeletal muscle cells (40%, P < 0.001). The endogenous expression of specific TFs may circumvent the additional requirement for TF reactivation and form a shortcut for cell fate conversion, suggesting a basic principle that could ease cell fate conversion. CONCLUSIONS: Our study demonstrates the first report of melanocyte-to-muscle conversion by small molecules, suggesting a novel strategy for muscle regeneration. Furthermore, skin is one of the tissues closely located to skeletal muscle, and therefore, our results provide a promising foundation for therapeutic chemical reprogramming in vivo treating skeletal muscle degenerative diseases.

Programmable A-to-Y base editing by fusing an adenine base editor with an N-methylpurine DNA glycosylase.

Here we developed an adenine transversion base editor, AYBE, for A-to-C and A-to-T transversion editing in mammalian cells by fusing an adenine base editor (ABE) with hypoxanthine excision protein N-methylpurine DNA glycosylase (MPG). We also engineered AYBE variants enabling targeted editing at genomic loci with higher transversion editing activity (up to 72% for A-to-C or A-to-T editing).

m6A reader HNRNPA2B1 destabilization of ATG4B regulates autophagic activity, proliferation and olaparib sensitivity in breast cancer.

N6-methyladenosine RNA (m6A) is the most extensive epigenetic modification in mRNA and influences tumor progression. However, the role of m6A regulators and specific mechanisms in breast cancer still need further study. Here, we investigated the significance of the m6A reader HNRNPA2B1 and explored its influence on autophagy and drug sensitivity in breast cancer. HNRNPA2B1 was selected by bioinformatics analysis, and its high expression level was identified in breast cancer tissues and cell lines. HNRNPA2B1 was related to poor prognosis. Downregulation of HNRNPA2B1 reduced proliferation, enhanced autophagic flux, and partially reversed de novo resistance to olaparib in breast cancer. ATG4B was determined by RIP and MeRIP assays as a downstream gene of HNRNPA2B1, by which recognized the m6A site in the 3'UTR. Overexpression of ATG4B rescued the malignancy driven by HNRNPA2B1 in breast cancer cells and increased the olaparib sensitivity. Our study revealed that the m6A reader HNRNPA2B1 mediated proliferation and autophagy in breast cancer cell lines by facilitating ATG4B mRNA decay and targeting HNRNPA2B1/m6A/ATG4B might enhance the olaparib sensitivity of breast cancer cells.

Midterm results of latent outflow tract obstruction in hypertrophic cardiomyopathy after septal myectomy: A propensity score-matched study.

BACKGROUND AND AIM OF THE STUDY: The midterm clinical outcomes of patients with latent left ventricular outflow tract (LVOT) obstruction who undergo septal myectomy are unclear. Therefore, this study aimed to evaluate the clinical outcomes of patients with latent LVOT obstruction who underwent septal myectomy. METHODS: We studied 34 patients with hypertrophic cardiomyopathy (HCM) and latent LVOT obstruction who underwent septal myectomy in 2011-2019 at Anzhen Hospital. After 2:1 propensity score matching, the study cohort included 34 patients with latent LVOT obstruction and 68 patients with resting LVOT obstruction. RESULTS: Compared to patients with resting LVOT obstruction, patients with latent LVOT obstruction had a thinner interventricular septal thickness (18.2 ± 3.2 mm vs. 20.4 ± 5.6 mm; p = .01), while the proportion of moderate or severe mitral regurgitation was significantly higher (26.5% vs. 5.9%; p = .003). Moreover, the proportion of mitral valve procedures (26.5% vs. 5.9%; p = .004) was significantly higher in patients with latent LVOT obstruction. However, there was no intergroup difference in cardiovascular death (5.9% vs. 1.5%, p = .26). Furthermore, the 5-year survival rates after sudden cardiac death (100.0% vs. 91.7%; p = .26) and cardiovascular death (95.5% vs. 89.0%; p = .32) were similar between HCM patients with latent versus resting LVOT obstruction. CONCLUSIONS: Midterm clinical outcomes were similar and excellent in a matched cohort of HCM patients with latent versus resting LVOT obstruction after septal myectomy.

Occludin is a target of Src kinase and promotes lipid secretion by binding to BTN1a1 and XOR.

Lipid droplets (LDs) have increasingly been recognized as an essential organelle for eukaryotes. Although the biochemistry of lipid synthesis and degradation is well characterized, the regulation of LD dynamics, including its formation, maintenance, and secretion, is poorly understood. Here, we report that mice lacking Occludin (Ocln) show defective lipid metabolism. We show that LDs were larger than normal along its biogenesis and secretion pathway in Ocln null mammary cells. This defect in LD size control did not result from abnormal lipid synthesis or degradation; rather, it was because of secretion failure during the lactation stage. We found that OCLN was located on the LD membrane and was bound to essential regulators of lipid secretion, including BTN1a1 and XOR, in a C-terminus-dependent manner. Finally, OCLN was a phosphorylation target of Src kinase, whose loss causes lactation failure. Together, we demonstrate that Ocln is a downstream target of Src kinase and promotes LD secretion by binding to BTN1a1 and XOR.

Controlling the Iron Migration Mechanism for the Cretaceous Sediment Color Variations in Sichuan Basin, China.

Cretaceous continental sediments in Sichuan Basin, China, have different colors, and the reasons for their formation are not determined. Based on mineralogical and geochemical characteristics, red beds and nonred beds in the Upper and Lower Cretaceous sedimentary strata in the western Sichuan Basin are described and tested in this study. The test and analysis of the mineral composition, element content, and iron speciation of mudstone samples with gray-green, gray, and red colors in Cangxi, Bailong, and Guankou formations found that the change in hematite content directly causes the color difference of samples. For red mudstone, the average chemical index of alteration, chemical index of weathering, weather eluviation index (Ba), Ca/(Mg*Al), and Al2O3/SiO2 index are 67.75, 79.94, 2.07, 0.26, and 0.26, respectively, indicating that chemical weathering is the most intense. The geochemical indexes corresponding to gray samples are 64.41, 74.91, 2.08, 0.19, and 0.24, respectively. Those corresponding to the gray-green samples are 62.30, 70.68, 2.17, 0.21, and 0.24, with the weakest chemical weathering. The ratio of Cu/Zn and the enrichment factor of V show that red and nonred bed samples are formed in weak oxidation and weak reduction environments, respectively. The red sample contains the highest content of hematite iron. The gray-green sample mainly represents paramagnetic ferrous in clay minerals. The geochemical contents of the gray sample's three iron elements are slightly different, mainly trivalent iron. The change in iron speciation content in different color samples shows that the Fe element forming hematite in red bed samples may come from the weathering of source rock and clay minerals subjected to secondary weathering. At present, it is confirmed that different colors of samples are related to different weathering degrees of source rocks, which can be related to hot, dry/humid climates. It is necessary to distinguish the climate type in combination with other indicators.

Summary of 20 tracheal intubation by anesthesiologists for patients with severe COVID-19 pneumonia: retrospective case series.

SARS-CoV-2 pandemic is announced and it is very important to share our experience to the critical care community in the early stage. Urgent intubation team was organized by anesthesiologists and was dispatched upon request. We have retrospectively reviewed medical charts of 20 critically ill patients with Covid-19 pneumonia who required tracheal intubation from February 17 to March 19 in Wuhan No.1 hospital, China. We collected their demographics, vital signs, blood gas analysis before and after tracheal intubation, and 7-day outcome after tracheal intubation. Out of 20 patients, 90% were over 60 years old and 15 were with at least one comorbidity. All meet the indication for tracheal intubation announced by treatment expert group. We had successfully intubated all patients using personal protective equipment without circulatory collapse during tracheal intubation. During the observational period, none of 17 anesthesiologists were infected. Although intubation improved SPO2, reduced PaCO2 and blood lactate, seven of 20 patients died within 7-days after tracheal intubation. Non-survivors showed significantly lower SPO2 and higher PaCO2 and blood lactate compared to survivors. For those who are anticipated to deteriorate severe pneumonia with poor prognosis, earlier respiratory support with tracheal intubation may be advised to improve outcome.

Coupling hydrophilic interaction chromatography materials with immobilized Fe3+ for phosphopeptide and glycopeptide enrichment and separation.

Simultaneous profiling of protein phosphorylation and glycosylation is very important to elucidate the bio-functions of these proteins. However, simultaneous enrichment of glyco- and phosphopeptides is the bottleneck in proteomics because of the low abundance of these species and ion suppression from non-modified peptides in mass spectrometry (MS). In this study, Fe3+ immobilized hydrophilic interaction chromatography (HILIC) materials (termed polySD-SiO2, recently reported in our lab) and polySD-SiO2 in the HILIC mode were employed for the simultaneous enrichment and subsequent separation of glyco- and phosphopeptides. The Fe3+ immobilized polySD-SiO2 could selectively enrich glycopeptides and phosphopeptides and the co-enriched peptides were further fractionated with polySD-SiO2 in the HILIC mode. With the established method, glyco- and phosphopeptides were well enriched and divided into two fractions even from tryptic digests of a-casein, fetuin and BSA at a molar ratio of 1 : 2 : 400. Application of the established method to HeLa cell lysate resulted in a total of 1903 phosphopeptides and 141 glycosylation sites. These results demonstrate that the established method could selectively and simultaneously enrich and fractionate glyco- and phosphopeptides from complex peptide mixtures.

Uncalibrated Visual Servoing for Underwater Vehicle Manipulator Systems with an Eye in Hand Configuration Camera.

This paper presents an uncalibrated visual servoing scheme for underwater vehicle manipulator systems (UVMSs) with an eye-in-hand camera under uncertainties. These uncertainties contain vision sensor parameters, UVMS kinematics and feature position information. At first, a linear separation approach is addressed to collect these uncertainties into vectors, and this approach can also be utilized in other free-floating based manipulator systems. Secondly, a novel nonlinear adaptive controller is proposed to achieve image error convergence by estimating these vectors, the gradient projection method is utilized to optimize the restoring moments. Thirdly, a high order disturbance observer is addressed to deal with time-varying disturbances, and the convergence of the image errors is proved under the Lyapunov theory. Finally, in order to illustrate the effectiveness of the proposed method, numerical simulations based on a 9 degrees of freedom (DOFs) UVMS with an eye-in-hand camera are conducted. In simulations, the UVMS is expected to track a circle trajectory on the image plane, meanwhile, time-varying disturbances are exerted on the system. The proposed scheme can achieve accurate and smooth tracking results during simulations.

Changes in Autophagy Levels in Rat Myocardium During Exercise Preconditioning-Initiated Cardioprotective Effects.

The role of autophagy in the cardioprotection conferred by ischemic preconditioning (IPC) has been well described. This study aimed to investigate the changes in autophagy levels during the cardioprotective effects initiated by exercise preconditioning (EP).Rats were randomly divided into 4 groups: group C (control), group EP, group EE (exhaustive exercise), and group EP + EE (EP pretreatment at 0.5 hours before EE). The EP protocol included 4 periods of 10 minutes of treadmill running each at 30 m/minute with intervening 10 minute periods of rest. Hematoxylin-basic fuchsin-picric acid (HBFP) staining and plasma levels of cardiac troponin I (cTnI) were used to evaluate the ischemia-hypoxia injury in rat myocardium. Alteration levels in several autophagy proteins in the left ventricular myocardium were analyzed by Western blot. The phasic alterations of autophagy levels during EP-initiated cardioprotective phase were also examined.Compared with group C, the ischemia-hypoxia positive areas and IOD value in HBFP-staining and cTnI plasma levels increased significantly in group EE. Compared with group EE, the ischemia-hypoxia injury was markedly attenuated in group EP + EE. Compared with group C, the LC3-II/LC3-I ratio, a marker of autophagosome formation, was reduced in group EE, but the LC3-II/LC3-I ratio remained unaltered in group EP + EE. Furthermore, the LC3-II/LC3-I ratio increased significantly at 2 hours during the cardioprotective phase after EP.These results suggest that the activated autophagy level during the EP-initiated cardioprotective phase may be partly involved in the cardioprotective effects by maintaining a normal autophagy basal level during the subsequent exhaustive exercise in rat myocardium.

Postoperative analgesia of Ropivacaine combined Dexmedetomidine for transversus abdominis plane block in laparoscopic surgery / 中国内镜杂志

Objective To observe the effect of postoperative analgesia of Ropivacaine combined Dexmedetomidine for transversus abdominis plane (TAP) block in laparoscopic surgery. Methods 60 patients underwent selective laparoscopic hernia repair were randomly divided into two groups: Ropivacine plus Dexmedetomidine group (Rpd group): Dexmedetomidine 1.0 μg/kg + 0.4% Ropivacine 30 ml was injected into bilateral TAP, and Ropivacine group (R group): saline 1 ml+0.4% Ropivacine 30 ml was injected into bilateral TAP. Blood pressure and heart rate on beginning of the surgery, 1 h during surgery and leaving the operating room, VAS of immediate postoperative period, 4 h, 6 h, 8 h, 12 h, 24 h after the surgery, analgesic duration and incidence of nausea and vomiting of the two groups were recorded. Results Two groups have no difference in blood pressure and heart rate in all time points (P > 0.05), VAS score of 6 h, 8 h, 12 h after surgery in Rpd group were lower than in R group (P < 0.05), and VAS score of postoperative period, 4 h, 24 h after the surgery had no difference between the two groups (P > 0.05). Sensory block duration in Rpd group was longer than in R group (P < 0.05). Incidence of nausea and vomiting of the two groups had no difference in two groups (P > 0.05). Conclusions Ropivacaine combined Dexmedetomidine for Transversus Abdominis Plane block in laparoscopy patients can significantly prolong the analgesic duration.

Postoperative analgesia of Ropivacaine combined Dexmedetomidine for transversus abdominis plane block in laparoscopic surgery / 中国内镜杂志

Objective To observe the effect of postoperative analgesia of Ropivacaine combined Dexmedetomidine for transversus abdominis plane (TAP) block in laparoscopic surgery. Methods 60 patients underwent selective laparoscopic hernia repair were randomly divided into two groups: Ropivacine plus Dexmedetomidine group (Rpd group): Dexmedetomidine 1.0 μg/kg + 0.4% Ropivacine 30 ml was injected into bilateral TAP, and Ropivacine group (R group): saline 1 ml+0.4% Ropivacine 30 ml was injected into bilateral TAP. Blood pressure and heart rate on beginning of the surgery, 1 h during surgery and leaving the operating room, VAS of immediate postoperative period, 4 h, 6 h, 8 h, 12 h, 24 h after the surgery, analgesic duration and incidence of nausea and vomiting of the two groups were recorded. Results Two groups have no difference in blood pressure and heart rate in all time points (P > 0.05), VAS score of 6 h, 8 h, 12 h after surgery in Rpd group were lower than in R group (P < 0.05), and VAS score of postoperative period, 4 h, 24 h after the surgery had no difference between the two groups (P > 0.05). Sensory block duration in Rpd group was longer than in R group (P < 0.05). Incidence of nausea and vomiting of the two groups had no difference in two groups (P > 0.05). Conclusions Ropivacaine combined Dexmedetomidine for Transversus Abdominis Plane block in laparoscopy patients can significantly prolong the analgesic duration.

[Effect of Dexmedetomidine Combined Electrical Stimulation on Coanitive Function of Patients Receiving Extracerebral Intervention].

OBJECTIVE: To explore the effect of dexmedetomidine combined electrical stimulation on cognitive function of neurosurgical diseases patients treated by extracerebral intervention. METHODS: Totally 122 patients with neurosurgical diseases who underwent selective intervention were randomly assigned to the observation group and the control group, 61 cases in each group. Patients in the control group recieved anesthesia by dexmedetomidine. Those in the observation group received electrical stimulation at Baihui (DU20), Yintang ( EX-HN3), and Neiguan (PC6) before dexmedetomidine anesthesia. The cognitive function of patients at preoperative day 1 and postoperative day 1 was respectively evaluated by Mini-Mental State Examinations (MMSE). Serum NSE, S-100ß, IL-1ß, IL-6, and TNF-α levels were detected in the two groups before intervention and immediately after intervention using ELISA. RESULTS: MMSE scores of two groups were significantly reduced at post-intervention day 1, as compared with one day before intervention. MMSE score of the observation group at post-intervention day 1 was (23.15 ± 1.87) points, significantly higher than that of the control group [ (19.34 ± 1.64) points , (P < 0.05)]. The postoperative cognitive dysfunction (POCD) incidence rate of the observation group was 16.4% (10/61), significantly lower than that of the control group [39.3% (24/61); P < 0.05]. Compared with before intervention, NSE and S-100ß protein levels, IL-1ß, IL-6 and α-TNF levels of the two groups increased (P < 0.05). Post-intervention NSE and S-100ß protein levels, IL-1ß, IL-6 and α-TNF levels were significantly lower in the observation group than in the control group (P < 0.05). CONCLUSION: Dexmedetomidine combied electrical stimulation could effectively prevent the occurrence of postoperative cognition, and reduce levels of NSA, S-100ß, IL-1ß, IL-6 and TNF-α.

[Protection of Transcutaneous Acupoint Electrical Stimulation for Brain Injury Undergoing Intervention: a Clinical Observation].

OBJECTIVE: To observe the effect of transcutaneous acupoint electrical stimulation (TAES) combined dexmedetomidine on hemodynamic of intracranial aneurysmal subarachnoid hemorrhage patients undergoing intervention, and their protection for brain Injury. METHODS: Totally 108 intracranial aneurysmal subarachnoid hemorrhage patients undergoing intervention were randomly assigned to the electroacupuncture (EA) group and the control group according to random digit table, 54 in each group. All patients were anesthetized with dexmedetomidine. Patients in the EA group were needled at bilateral Neiguan (PC6), Lieque (LU7), and Yunmen (LU2). Parameter setting was as follows: The dilatational wave at 1. 5 Hz, strength 2 - 4 mA, 30 min. Systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), and heart rate (HR) were compared between the two groups immediately after entry into the room (T0), after administration (T1), intubating (T2), resuscitation (T3), extubation (T4), and leaving the operating room (T5). Levels of S100ß protein (S100ß) and neuron specific enolase (NSE) were compared between the two groups at T0, immediately after surgery (T6), 6 h after operation (T7), 12 h after operation (T8), and 24 h after operation (T9). RESULTS: Compared with the same group at T0, SBP, DBP, MAP, and HR were significantly reduced in the two groups at T1-T5(P <0. 05), serum levels of S100ß and NSE in the two groups were significantly increased at T6-T9 (P<0. 05). Compared with the control group at T1 - T5, SBP, DBP, MAP, and HR decreased in the EA group (P <0. 05). Compared with the control group at T6-T9, serum levels of S100ß and NSE decreased in the EA group (P <0. 05). CONCLUSION: TAES combined dexmedetomidine could effectively maintain stable hemodynamics of intracranial aneurysmal subarachnoid hemorrhage patients undergoing intervention, and regulate their serum levels of S100ß and NSE.

A modified chordal transfer technique to treat anterior mitral leaflet prolapse.

Chordal transfer from the intact posterior mitral leaflet (PML) to the anterior mitral leaflet (AML) is an effective way to correct anterior leaflet prolapse and provides good long-term results. However, it is difficult to determine the accurate segment of the PML which needs to be transferred and the suture point of the leaflets. We describe a modified technique to determine the correct segment that needs to be transferred to effectively correct AMLs with elongated or ruptured chordae. This technique renders performing chordal transfer easier and more accurate.