Down-regulation of the Smad signaling by circZBTB46 via the Smad2-PDLIM5 axis to inhibit type I collagen expression / 老年心脏病学杂志（英文版）

BACKGROUND@#Abnormal type I collagen (COL1) expression is associated with the development of many cardiovascular diseases. The TGF-beta/Smad signaling pathway and circRNAs have been shown to regulate COL1 gene expression, but the underlying molecular mechanisms are still not fully understood.@*METHODS@#Gain- and loss-of-function experiments were prformed to study the effect of circZBTB46 on the expression of alpha 2 chain of type I collagen (COL1A2). Co-immunoprecipitation assay was performed to observe the interaction between two proteins. RNA immunoprecipitation assay and biotin pull-down assay were performed to observe the interaction of circZBTB46 with PDLIM5.@*RESULTS@#In this study, we investigated the role of circZBTB46 in regulating COL1A2 expression in human vascular smooth muscle cells (VSMCs). We found that circZBTB46 is expressed in VSMCs and that TGF-beta inhibits circZBTB46 formation by downregulating KLF4 expression through activation of the Smad signaling pathway. CircZBTB46 inhibits the expression of COL1A2 induced by TGF-beta. Mechanistically, circZBTB46 mediates the interaction between Smad2 and PDLIM5, resulting in the inhibition of Smad signaling and the subsequent downregulation of COL1A2 expression. Furthermore, we found that the expression of TGF-beta and COL1A2 is decreased, while circZBTB46 expression is increased in human abdominal aortic aneurysm tissues, indicating that circZBTB46-mediated regulation of TGF-beta/Smad signaling and COL1A2 synthesis in VSMCs plays a crucial role in vascular homeostasis and aneurysm development.@*CONCLUSIONS@#CircZBTB46 was identified as a novel inhibitor of COL1 synthesis in VSMCs, highlighting the importance of circZBTB46 and PDLIM5 in regulating TGF-beta/Smad signaling and COL1A2 expression.

Mechanism of Xiaojinwan Against Breast Cancer Bone Metastasis： An Exploration Based on Bioinformatic Analysis Combined with Cell Experiments / 中国实验方剂学杂志

Objective:To explore the mechanism of Xiaojinwan in treating breast cancer bone metastases through cell experiments and bioinformatic analysis. Method:The inhibitory effect of Xiaojinwan on MCF-7 cell viability was detected by cell counting kit-8 （CCK-8） assay. The key components and targets responsible for Xiaojinwan in inhibiting breast cancer bone metastases were predicted by network pharmacology and molecular docking. The active components and targets of Xiaojinwan were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCSMP） and SwissTarget Prediction， and the breast cancer bone metastases-related targets from GeneCards and DisGeNET. The results were imported into STRING for constructing a protein-protein interaction （PPI） network， followed by Gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis using DAVID. A network of the active components of Xiaojinwan-breast cancer bone metastases-related targets-pathways was constructed using Cytoscape 3.7.2. AutoDock 4 was employed for molecular docking. The protein expression levels of matrix metallopmteinase-9 （MMP-9）， hypoxia-inducible factor 1<italic>α </italic>（HIF1A）， and androgen receptor （AR） were assayed by Western blot. Result:Xiaojinwan inhibited the viability of MCF-7 cells and acted on breast cancer bone metastases through such processes as redox and protein autophosphorylation. KEGG enrichment analysis showed that HIF-1， vascular endothelial growth factor （VEGF） and phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） pathways were involved. As verified by molecular docking， the active components such as eucalyptin stably bound to AR and MMP-9. Western blot indicated that Xiaojinwan dose-dependently inhibited the expression of MMP-9 and HIF1A proteins in MCF-7 cells. Conclusion:Xiaojinwan acts on AR and MMP-9 through HIF， VEGF and other related signaling pathways， thereby improving hypoxia in tumor microenvironment， inhibiting angiogenesis， and reducing cell invasion and viability.

SIX1 Promotes Proliferation and Invasion of Breast Cancer Cells / 中国生物化学与分子生物学报

Sine oculis homeobox homolog 1 (SIX1) is involved in the regulation of many kinds of tumors and plays an important role in the occurrence and development of breast cancer, but the exact mechanism remains to be explored. In this study, we analyzed the expression of SIX1 in breast cancer, and investigated the role of SIX1 in the proliferation and invasion of breast cancer cells. TCGA database and GEPIA2 showed that the expression of SIX1 in breast cancer was significantly higher than that in normal tissues, which was confirmed in different molecular subtypes of breast cancer, including Basal-like, HER2, Luminal A and Luminal B (P < 0. 05). Besides, the analysis of HPA database also showed that the expression of SIX1 was significantly upregulated in breast cancer, and the difference was statistically significant (P < 0. 001). After interfering with the expression of SIX1 in the breast cancer cell line MDA-MB-231 and overexpressing SIX1 in MCF-7, the growth curve and EdU results showed that the proliferation of breast cancer cells was inhibited after knocking down SIX1, while overexpression of SIX1 could promote the proliferation ability (the growth curve assays: P < 0. 05; EdU assays: P < 0. 001). Besides, Transwell assays showed that SIX1 could enhance the invasion ability of breast cancer cells (P < 0. 001). In TCGA database we defined the high and low expression populations according to the upper and lower quartiles of SIX1 gene expression, and differentially expressed genes were found to be associated with metabolism and stem cell regulatory pathways. For further confirmation, high-throughput RNA deep sequencing (RNA-seq) was conducted using SIX1-knockdown cells, and analysis also showed that SIX1 was closely related to metabolism, stem cell regulation and EMT pathways. We selected several representative genes, MYC, SNAI2 and EGFR, to examine their associations with SIX1 expression and prognosis in patients with clinical breast cancer using published GEO datasets and KM-plotter, we found that SIX1 was positively correlated with the expression of MYC, SNAI2 and EGFR, and the high expression of SIX1, MYC, SNAI2 and EGFR is not conducive to the survival of breast cancer patients. GCBI, GeneMANIA and String online tools were conducted to predict the associated genes, lncRNA and miRNA of SIX1. Collectively, our study initially revealed the role of SIX1 in breast cancer and its regulatory mechanism, providing new insights for further studies.

The metabolic networks of ferroptosis and links to lung diseases / 生理学报

Ferroptosis is a newly discovered non-apoptotic form of regulated cell death driven by iron-dependent lipid peroxidation. The present studies have shown that many metabolic processes and homeostasis are affected by ferroptosis. It is related to many lung diseases, including acute lung injury, chronic obstructive pulmonary disease and pulmonary fibrosis, etc. Currently, the research on ferroptosis is still in its infancy. Previous studies have confirmed that ferroptosis is regulated by a variety of genes, and the mechanism is complex, mainly involving iron homeostasis and lipid peroxidation metabolism. This review summarizes some regulation networks of metabolic processes associated with ferroptosis and discusses the roles of ferroptosis in the pathophysiological progression of many lung diseases. We expected to provide new ideas and references for the treatment of these diseases.

The construction and application of online open courses in Chinese colleges and universities--Taking the physiology course at Central South University of China as an example / 生理学报

Massive open online course (MOOC) is a new learning model, which integrates the progress of novel educational concepts and the breakthrough of information technology. MOOC uses new web-based tools and online-environments to deliver knowledge education and lecture classes in a new paradigm. In this paper, we firstly reviewed the achievements through four stages of the construction and development of online courses of physiology in China in the past 20 years. Then, taking the physiology MOOC at Central South University of China as an example, we introduced the specific practices and experiences to construct the online physiological open course, including the online open course-based offline and online flipped classroom teaching practice. Finally, we discussed several important issues during the construction and application of online open courses, aiming to provide practical information for other universities.

C5-6 nerve root block technique for postoperative analgesia of shoulder arthroscope: a randomized controlled trial / 北京大学学报(医学版)

OBJECTIVE@#To compare the effects of ultrasound-guided interscalene brachial plexus block and C5-6 nerve root block for analgesia after shoulder arthroscopy.@*METHODS@#In the study, 40 patients of ASA I-II were selected for elective general anesthesia to repair the shoulder ligament rupture in Peking University Third Hospital, who were randomly divided into two groups, respectively for the intermuscular brachial plexus block group (group I) and C5-6 nerve root block group (group C), n=20. The forty patients underwent ultrasound-guided brachial plexus block or C5-6 nerve root block before general anesthesia. Group I: 0.2% ropivacaine 10 mL was injected into brachial plexus intermuscular approach; Group C: 0.2% ropivacaine 10 mL was injected around the nerve roots of C5 and C6, and the ultrasound images showed that the liquid wrapped nerve roots. The time of sensory and motor block after puncture, operation time, the time of postoperative analgesia, numerical rating scale (NRS) scores at 1, 6, 12, and 24 h postoperatively and the finger movements were recorded. The adverse drug reactions and the patient satisfaction were recorded. The primary end point was the study of shoulder rest and movement pain in the patients with postoperative nerve blockage; the secondary end point was the patient's limb movements and thepatient satisfaction.@*RESULTS@#The duration of analgesia was (571.50±70.11) min in group I and (615.60±112.15) min in group C, and there was no difference between the two groups (P>0.05). The static and dynamic NRS scores at 1, 6, and 12 h in group C were lower than those in group I (P<0.05). There was no difference in static and dynamic NRS scores between the two groups during 24 hours (P>0.05). There was a significant difference in grade of muscle strength between group C [5(4,5)] and group I [4(2,4)] in the patients with nerve block hind limb (P<0.01), and there were significant differences between the two groups' sensation in the radial nerve group C [1(0,2)] and group I [2(1,2)], the median nerve group C [0(0,2)] and group I [2(1,2)], and the ulnar nerves group C [0(0,1)] and group I [1(1,2)] (P<0.01). There was no statistical difference between the two groups in the sencation of the shoulder, group C 2(1,2) and group I 2(1,2) , P>0.05. Compared with group I 8(6,9), group C 9(8,10) was a significant difference in satisfaction (P<0.01).@*CONCLUSION@#Interscalene brachial plexus block and C5-6 nerve root block could satisfy the needs of analgesia after shoulder arthroscopy, but C5-6 nerve root blockage does not limit the limb activity, the numbness is less, and the patient's satisfaction is higher.

Rationality evaluation of predisone dosage in the treatement of SLE with different clinical scoring systems / 中华疾病控制杂志

Objective To evaluate the rationality of predisone dosage in the treatement of SLE with different clinical scoring systems. Methods The clinical data of 51 newly diagnosed patients with systemic lupus erythematosus(SLE) were collected, and disease activity was assessed by SLEDAI- 2000, BILAG- 2004 and SLAM-R scoring systems. The correlations between SLEDAI- 2000, BILAG- 2004 , SLAM-R scores and prednisone dosage were analyzed. Results BILAG-2004 score showed the best correlation with the dosage of prednision (r=0.827, P<0.001) .SLAM score showed the worst correlation with the dosage of prednision (r=0.512, P<0.001). Different treatment choice was associated with different organ/system involvement. Conclusions BILAG- 2004 , SLEDAI- 2000 and SLAM-R scoring systems all can be used as references for the prednisone dosage slection.It is suggested that the BILAG- 2004 scoring system is more reliable and comprehensive in clinical practice．

Role of neuropeptide substance P and the bone morphogenetic protein signaling pathway in osteogenic differentiation of ST2 cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study aimed to investigate the role and mechanism of neuropeptide substance P (SP) in ST2 cell (bone mesenchymal stem cells of mice) osteogenic differentiation to provide a basis for the treatment of temporomandibular joint osteoarthritis.</p><p><b>METHODS</b>Third-generation ST2 cells were cultured with different concentrations of SP (0, 10⁻¹⁰, 10⁻⁸, 10⁻⁶, and 10⁻⁵ mol·L⁻¹). After 24, 48, and 72 h, cell proliferation was detected by CCK-8. The ST2 cells were cultured with 10⁻⁶ mol·L⁻¹ SP for 1, 3, 5, and 7 days. Subsequently, the expression of alkaline phosphatase (ALP), collagen typeⅠ(CollaⅠ), and osteocalcin (OCN) in the culture supernatant was tested by enzyme-linked immunosorbent assay (ELISA). ALP activity was detected by immunofluorescence staining. The ST2 cells were cultured with SP, Noggin (inhibitor of the bone morphogenetic protein signaling pathway), SP+Noggin, and 2% fetal bovine serum, respectively. Finally, the expression of ALP, CollaⅠ, and OCN in the culture supernatant was tested by ELISA.</p><p><b>RESULTS</b>CCK-8 showed that the effect of cell proliferation was most obvious when the SP concentration was 10⁻⁶ mol·L⁻¹ (P<0.01). The ELISA results demonstrated that ALP expression significantly increased at day 5 compared with that in the control group (P<0.01), whereas the expression of CollaⅠand OCN significantly increased at day 7 (P<0.05). Immunofluorescence results showed that ALP activity was strongest at day 5. The expression of ALP, CollaⅠ, and OCN decreased after Noggin addition (P<0.05).</p><p><b>CONCLUSIONS</b>SP can promote the proliferation and osteogenic differentiation of ST2 cells, and the bone morphogenetic protein signaling pathway may be involved in this process.</p>

Mechanical properties of human enamel based on resonant ultrasound spectroscopy / 医用生物力学

Objective To investigate the mechanical properties of human enamel based on resonant ultrasound spectroscopy (RUS). Methods The rectangular parallelepiped specimens of human enamel were processed. The theoretical resonant frequencies of specimens were estimated and paired with the experimental resonant frequencies measured from RUS experiments. An iterative procedure was used to adjust elastic constants of enamel until the theoretical frequencies corresponded to the experimental frequencies based on minimum mean-squared error criterion. In addition, elastic modulus, shear modulus and Poisson’s ratio were calculated respectively. Results The elastic modulus, shear modulus and Poisson’s ratio ranged from 61.52 to 80.46 GPa, 21.51 to 51.86 GPa and 0.18 to 0.43, respectively. Eliminating the effect of large specimen variances, the average of elastic modulus, shear modulus and Poisson’s ratio was 72.84 GPa, 31.94 GPa and 0.27, respectively. Conclusions RUS performs a feasibility of measuring the mechanical properties of human enamel with repeatable and nondestructive advantages. All the elastic constants and mechanical parameters can be estimated through a signal experiment. The results provide references for the development of biomimetic dental restoration materials.

Mechanical properties of human enamel based on resonant ultrasound spectroscopy / 医用生物力学

Objective To investigate the mechanical properties of human enamel based on resonant ultrasound spectroscopy (RUS).Methods The rectangular parallelepiped specimens of human enamel were processed.The theoretical resonant frequencies of specimens were estimated and paired with the experimental resonant fre quencies measured from RUS experiments.An iterative procedure was used to adjust elastic constants of enamel until the theoretical frequencies corresponded to the experimental frequencies based on minimum mean-squared error criterion.In addition,elastic modulus,shear modulus and Poisson's ratio were calculated respectively.Results The elastic modulus,shear modulus and Poisson's ratio ranged from 61.52 to 80.46 GPa,21.51 to 51.86 GPa and 0.18 to 0.43,respectively.Eliminating the effect of large specimen variances,the average of elastic modulus,shear modulus and Poisson's ratio was 72.84 GPa,31.94 GPa and 0.27,respectively.Conclu sions RUS performs a feasibility of measuring the mechanical properties of human enamel with repeatable and nondestructive advantages.All the elastic constants and mechanical parameters can be estimated through a sig nal experiment.The results provide references for the development of biomimetic dental restoration materials.

Metabotropic glutamate receptor 8 activation promotes the apoptosis of lung carcinoma A549 cells in vitro / 生理学报

This study aims to detect the expression of metabotropic glutamate receptors (mGluRs) in lung carcinoma A549 cells, and to investigate the effects of mGluR8 and mGluR4 activation on the growth of A549 cells in vitro. The mRNA expression levels of the 8 subtypes of mGluRs in A549 cells were determined by real-time PCR. Immunohistochemistry was used to analyze the protein expression of mGluR4 and mGluR8 in A549 cells and lung tissue sections obtained from lung adenocarcinoma patients. To observe the effects of mGluR8 and mGluR4 activation on the growth of A549 cells, the cultured cells were treated with (S)-3,4-DCPG (an agonist of mGluR8) and VU0155041 (an agonist of mGluR4), respectively, and then the cell viability was analyzed by CCK-8 kit, the percentage of DNA synthesis was detected by EdU incorporation, and the apoptosis of the cells was measured by hoechst 33258 staining and flow cytometry. The results showed that there were low expressions of mGluR1, mGluR5, mGluR6, mGluR7 mRNA, no expression of mGluR2 and mGluR3 mRNA, and high expressions of mGluR8 and mGluR4 mRNA in A549 cells. Accordingly, there were also mGluR4 and mGluR8 protein expressions in the A549 cells and the lung adenocarcinoma tissue sections. VU0155041 had no effect on the growth of A549 cells, but (S)-3,4-DCPG significantly decreased the cells' growth in a dose-dependent manner and increased the apoptosis of the cells. The results revealed a role of mGluR8 in the growth and apoptosis of A549 cells and suggested a potential target for clinical treatment of lung cancer.

A brief analysis of taking off the needle slowly / 中国针灸

The ancients highlight taking off the needle slowly. From aspects of proper timing of withdrawing the needle, the safe treatment, treating and keeping the spirit, dealing with the acupuncture accidents, reinforcing and reducing manipulation and strengthening the post-needling sensation, etc. advantages of taking off the needle slowly are described in this paper. So the taking off the needle slowly is worthwhile in clinic.

Bioinformatics of mouse uteroglobin binding protein and its polyclonal antibody preparation / 生理学报

To prepare anti-mouse uteroglobin binding protein (mUGBP) polyclonal antibody, two polypeptides were synthesized based on the bioinformatics analysis of mUGBP, and New Zealand white rabbits were immunized separately with each peptide coupled with keyhole limpet hemocyanin (KLH). The data indicate that a 13-amino acid polypeptide (positions 221st-233rd) was able to generate anti-peptide antibodies. The titer of the antisera detected with ELISA was 1:10(8). The antisera were then purified with immuno-affinity chromatography to obtain antibodies. Western blot analysis of mUGBP expressed as a fusion protein with a green fluorescent protein (GFP) was performed on the cell lysates of COS-1 cells with the purified antisera, suggesting that the antisera specifically recognized UGBP. By immunohistochemistry and indirect immunofluorescence analysis, we examined the expression of UGBP in the lung tissues from a patient undergoing surgical lung resection for a tumor and from normal mouse lung tissue, and found for the first time that UGBP protein was widely expressed in both mouse and human lung tissue with the most abundant expression in bronchial epithelial cells. These results suggest that the antigen epitopes of mUGBP are well predicted by using bioinformatics analysis. We have obtained anti-mUGBP polyclonal antibody, which will be useful for further investigation.

Activation of mGluRI in neutrophils promotes the adherence of neutrophils to endothelial cells / 生理学报

L-glutamate (Glu) is an excitatory neurotransmitter in the mammalian central nervous system. Relatively much attention has been paid to functional expression of Glu signaling molecules in peripheral tissues very recently. The present study tested the hypothesis that the activation of group I metabotropic glutamate receptor (mGluRI) in neutrophils stimulated neutrophils adherence to endothelial cells by increasing the surface expression of certain adhesion molecules. Peripheral blood was obtained by venipuncture from healthy donors, and the neutrophils were isolated by Ficoll-Hypaque gradient centrifugation. Neutrophils floating into DMEM/F12 culture medium containing 10% fetal bovine serum were then used immediately. Immunocytochemistry and real-time quantitative RT-PCR were used to detect the expression of mGluRI (mGluR1 and mGluR5) in neutrophils. The adherence of neutrophils to cultured human normal umbilical vein endothelial cells (HUVE-12) was measured by the colorimetric method. Cell surface expression of adhesion molecule CD11a in the neutrophils was determined by flow cytometry. Immunocytochemistry and real-time quantitative RT-PCR showed that mGluR1 and mGluR5 were constitutively expressed in neutrophils. Application of mGluRI agonist S-3,5-dihydroxyphenylglycine (S-DHPG) (1x10(-8)-1x10(-6) mol/L) showed a dose-dependent stimulatory effect on the adherence of neutrophils to HUVE-12 (P<0.05 or P<0.01), with a maximum effect at 1x10(-6) mol/L (P<0.01). Incubations as short as 30 min were sufficient to induce increased adherence after the beginning of S-DHPG treatment. Following time extension (0.5-5 h), S-DHPG (1x10(-6) mol/L) increased the rate of neutrophils adhesion to HUVE-12 with a maximum effect at 0.5 h (P<0.01). However, a time-dependent effect of S-DHPG on the rate of neutrophils adhesion to HUVE-12 was not observed during the experimental period. 1x10(-6) mol/L of S-DHPG also induced an increased surface expression of adhesion molecule CD11a (P<0.01) when neutrophils were preincubated with 1x10(-6) mol/L of S-DHPG for 1 h. Furthermore, the specific mGluRI antagonist (RS)-alpha-methyl-4-carboxyphenylglycine ((+/-)-MCPG, 0.5 mmol/L) significantly abolished the stimulatory effect of S-DHPG (1x10(-6) mol/L) on the adherence of neutrophils to HUVE-12 (P<0.01). These results suggest that the activation of mGluRI in neutrophils results in increased adhesion molecule CD11a expression and thereby promotes the adherence of neutrophils to endothelial cells.

Protective effect of low concentration endothelin-1 on the reactive oxygen-induced inhibition of pulmonary surfactant lipid synthesis / 生理学报

The effects of endothelin-1 (ET-1) at low concentration (1-100 pmol/L) on the reactive oxygen-induced inhibition of both pulmonary surfactant (PS) lipid synthesis and the activity of CTP: phosphorylcholine cytidylyltransferase (CCT), a rate-limiting enzyme in biosynthesis of phosphoatidylcholine (PC), were studied in cultured lung explants without serum. The xanthine-xanthine oxidase superoxide anion generating system decreased (3)H-choline incorporation into PC in a dose-dependent manner in cultured lung explants. ET-1 reduced both the reactive oxygen-induced decrease in (3)H-choline incorporation and the increase in malondialdehyde (MDA) content of lung tissues, but did not change the levels of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and the total antioxidant capability in the lung explants. ET-1 enhanced microsomal CCT activity of the lung tissues, while it decreased cytosolic CCT activity of lung tissues. ET-1 also prevented the inhibitive effect of reactive oxygen on microsomal CCT activity in the lung explants. These results suggest that ET-1 at low concentration can protect the microsomal CCT activity and reduce the inhibition of PS lipid synthesis induced by oxidant lung injury. The protective mechanism of ET-1 is not relative to the pulmonary endogenous antioxidant defense system.