The RCAN1.4-calcineurin/NFAT signaling pathway is essential for hypoxic adaption of intervertebral discs

Huang Bao,He Yongqing,Li Shengwen,Wei Xiaoan,Liu Junhui,Shan Zhi,Huang Yue,Chen Jiang,Zhao Fengdong 생화학분자생물학회 2020 Experimental and molecular medicine Vol.52 No.-

Calcipressin-1, also known as regulator of calcineurin 1 (RCAN1), can specifically bind calcineurin at or near the calcineurin A catalytic domain and downregulate calcineurin activity. However, whether RCAN1 affects the hypoxic intervertebral disc (IVD) phenotype through the calcineurin/NFAT signaling pathway remains unclear. First, we confirmed the characteristics of the degenerative nucleus pulposus (NP) by H&E, safranin O/fast green and Alcian blue staining, and detected increased RCAN1 levels in the degenerative NP by immunohistochemistry. Then, we demonstrated that the protein level of RCAN1.4 was higher than that of RCAN1.1 and progressively elevated from the control group to the Pfirrmann grade V group. In vitro, both hypoxia (1% O2) and overexpression of HIF-1α reduced the protein level of RCAN1.4 in rat NP cells in a dose- and time-dependent manner. We further found that miRNA-124, through a nondegradative pathway (without the proteasome or lysosome), suppressed the expression of RCAN1.4. As expected, calcineurin in NP cells was activated and primarily promoted nuclear translocation of NFATc1 under hypoxia or RCAN1.4 siRNA transfection. Furthermore, SOX9, type II collagen and MMP13 were elevated under hypoxia, RCAN1.4 siRNA transfection or NFATc1 overexpression. Using chromatin immunoprecipitation (ChIP) and a luciferase reporter assay (with mutation), we clarified that NFATc1 increasingly bound the SOX9 promotor region (bp −367~−357). Interaction of HIF-1α and NFATc1 promoted MMP13 transcription. Finally, we found that FK506 reversed hypoxia-induced activation of the calcineurin/NFAT signaling pathway in NP cells and an ex vivo model. Together, these findings show that the RCAN1.4-calcineurin/ NFAT signaling pathway has a vital role in the hypoxic phenotype of NP cells. RCAN1.4 might be a therapeutic target for degenerative disc diseases.

Novel DNAH1 Mutation Loci Lead to Multiple Morphological Abnormalities of the Sperm Flagella and Literature Review

Zhuang Bao-Jun,Xu Su-Yun,Dong Liang,Zhang Pei-Hai,Zhuang Bao-Lin,Huang Xiao-Peng,Li Guang-Sen,You Yao-Dong,Chen Di'Ang,Yu Xu-Jun,Chang De-Gui 대한남성과학회 2022 The World Journal of Men's Health Vol.40 No.4

The protein encoded by dynein axonemal heavy chain 1 (DNAH1) is a part of dynein, which regulates the function of cilia and sperm flagella. The mutant of DNAH1 causes the deletion of inner dynein arm 3 in the flagellum, leading to multiple morphological abnormalities of the sperm flagella (MMAF) and severe asthenozoospermia. However, instead of asthenozoospermia and MMAF, the result caused by the mutation of DNAH1 remains unknown. Here we report a male infertility patient with severe asthenozoospermia and teratozoospermia. We found two heterozygous mutations in DNAH1 (c.6912C>A and c.7076G>T) and which were reported to be associated with MMAF for the first time. We next collected and analyzed 65 cases of DNAH1 mutation and found that the proportion of short flagella is the largest, while the bent flagella account for the smallest, and the incidence of head deformity is not high in the sperm of these patients. Finally, we also analyzed 31 DNAH1 mutation patients who were treated with intracytoplasmic sperm injection (ICSI) and achieved beneficial outcomes. We hope our research will be helpful in the diagnosis and treatment of male infertility caused by DNAH1 mutation.

Local Extrema Refinement Based Tensor Product Model Transformation Controller Design with Vary Input Methods

Bao Shi,Guoliang Zhao,Sharina Huang 제어·로봇·시스템학회 2022 International Journal of Control, Automation, and Vol.20 No.4

Tensor product model transformation could find a convex polytope representation similar to the TakagiSugeno (TS) fuzzy model from the given quasi-linear variable parameter (quasi-LPV) system model. TP models have proposed many convex hulls manipulated methods in previous studies, but these convex hulls are designed based on classical sampling methods, and local extrema refinement methods are often ignored by these classical sampling methods. Therefore, this paper proposes a convex hull manipulate method by adding local extremum to refine the partition of the entry functions, and the method is proposed based on vary input quasi-LPV state space models. First, the TP model transformation is extended by linearizing the weighting functions that is obtained via different ways of substituted entry functions, the given TS fuzzy model is converted into an alternative TS fuzzy model, and the converted TS fuzzy models are presented with different number of inputs. The difficulty level of controller design is reduced by changing the antecedents of the fuzzy sets, so the optimized control can be achieved via linear matrix inequalities (LMIs). Secondly, the local extrema refinement strategy is used to implement the TP model transformation, the manipulated method of convex hull is expanded. Finally, the GOOGOL’s twowheeled self-balancing robot model is employed as the controlled object, and the tracking controller is attached for verification. At the same time, different external disturbances are considered to be added to the left and right wheels of the robot. The simulation results show that, by combining the state-space model with vary input variables and the local extrema refinement strategy, the designed controller achieved better control performance.

Comparative study on dynamic properties of argillaceous siltstone and its grouting-reinforced body

Huang, Ming,Xu, Chao-Shui,Zhan, Jin-Wu,Wang, Jun-Bao Techno-Press 2017 Geomechanics & engineering Vol.13 No.2

A comparison study is made between the dynamic properties of an argillaceous siltstone and its grouting-reinforced body. The purpose is to investigate how grout injection can help repair broken soft rocks. A slightly weathered argillaceous siltstone is selected, and part of the siltstone is mechanically crushed and cemented with Portland cement to simulate the grouting-reinforced body. Core specimens with the size of $50mm{\times}38mm$ are prepared from the original rock and the grouting-reinforced body. Impact tests on these samples are then carried out using a Split Hopkinson Pressure Bar (SHPB) apparatus. Failure patterns are analyzed and geotechnical parameters of the specimens are estimated. Based on the experimental results, for the grouting-reinforced body, its shock resistance is poorer than that of the original rock, and most cracks happen in the cementation boundaries between the cement mortar and the original rock particles. It was observed that the grouting-reinforced body ends up with more fragmented residues, most of them have larger fractal dimensions, and its dynamic strength is generally lower. The mass ratio of broken rocks to cement has a significant effect on its dynamic properties and there is an optimal ratio that the maximum dynamic peak strength can be achieved. The dynamic strain-softening behavior of the grouting-reinforced body is more significant compared with that of the original rock. Both the time dependent damage model and the modified overstress damage model are equally applicable to the original rock, but the former performs much better compared with the latter for the grouting-reinforced body. In addition, it was also shown that water content and impact velocity both have significant effect on dynamic properties of the original rock and its grouting-reinforced body. Higher water content leads to more small broken rock pieces, larger fractal dimensions, lower dynamic peak strength and smaller elastic modulus. However, the water content plays a minor role in fractal dimensions when the impact velocity is beyond a certain value. Higher impact loading rate leads to higher degree of fragmentation and larger fractal dimensions both in argillaceous siltstone and its grouting-reinforced body. These results provide a sound basis for the quantitative evaluation on how cement grouting can contribute to the repair of broken soft rocks.

Laboratory evaluation of differential attraction of Culex pipiens pallens to the volatiles of flowers, fruits, and seed pods

Bao-Ting Yu,Shu-Qi Huang,Yan-Mei Ding,Hatem Fouad,Hong-Jie Li,Jian-Chu Mo 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol.20 No.4

The attraction of Culex pipiens pallens Coquillett to various potential sugar sources in their habitats remains unknown. In the present study, we evaluated the attraction of Cx. pipiens pallens to the volatiles of different kinds of flowers, fruits, and seed pods by multiple choice tests under laboratory conditions. Nine of the 21 tested fresh tissues of flowers were attractive to females and six of them were attractive to males. The volatiles of six flowers (Ligustrum quihoui, Abelia chinensis, Ophiopogon japonicus, Phaseolus vulgaris, Osmanthus fragrans, and Nerium indicum) were attractive to both female and male Cx. pipiens pallens, and the flowers of Torenia fournieri and Lavandula angustifolia were potentially repellent to female and male mosquitoes, respectively. Besides, the volatiles of three fruits and seed pods (Broussonetia papyrifera, Amygdalus persica, and Cucumis melo) were attractive to both female and male Cx. pipiens pallens while the seed pods of Ficus carica and Pyrus bretschneideri were only attractive to females and males, respectively. This study provides valuable information on developing novel control methods against Cx. pipiens pallens in China.

CT-Guided Microcoil Localization of Small Peripheral Pulmonary Nodules to Direct Video-Assisted Thoracoscopic Resection without the Aid of Intraoperative Fluoroscopy

Huang Zhen-guo,Wang Cun-li,Sun Hong-liang,Li Chuan-dong,Gao Bao-xiang,Chen He,Yang Min-xing 대한영상의학회 2021 Korean Journal of Radiology Vol.22 No.7

Objective: To evaluate the feasibility, safety, and effectiveness of CT-guided microcoil localization of solitary pulmonary nodules (SPNs) for guiding video-assisted thoracoscopic surgery (VATS). Materials and Methods: Between June 2016 and October 2019, 454 consecutive patients with 501 SPNs who received CTguided microcoil localization before VATS in our institution were enrolled. The diameter of the nodules was 0.93 ± 0.49 cm, and the shortest distance from the nodules to the pleura was 1.41 ± 0.95 cm. The distal end of the microcoil was placed less than 1 cm away from the nodule, and the proximal end was placed outside the visceral pleura. VATS was performed under the guidance of implanted microcoils without the aid of intraoperative fluoroscopy. Results: All 501 nodules were marked with microcoils. The time required for microcoil localization was 12.8 ± 5.2 minutes. Microcoil localization-related complications occurred in 179 cases (39.4%). None of the complications required treatment. A total of 463 nodules were successfully resected under the guidance of implanted microcoils. VATS revealed 38 patients with dislocated microcoils, of which 28 underwent wedge resection (21 cases under the guidance of the bleeding points of pleural puncture, 7 cases through palpation), 5 underwent direct lobectomy, and the remaining 5 underwent a conversion to thoracotomy. In 4 cases, a portion of the microcoil remained in the lung parenchyma. Conclusion: CT-guided microcoil localization of SPNs is safe and reliable. Marking the nodule and pleura simultaneously with microcoils can effectively guide the resection of SPNs using VATS without the aid of intraoperative fluoroscopy.

PCSK9 regulates myocardial ischemia–reperfusion injury through parkin/pink1-mediated autophagy pathway

Huang Guangwei,Bao Hailong,Zhan Peng,Lu Xiyang,Duan Zonggang,Xiong Xinlin,Lin Muzhi,Wang Bing,An Hongxin,Xiahou Luanda,Zhou Haiyan,Luo Zhenhua,Li Wei 대한독성 유전단백체 학회 2024 Molecular & cellular toxicology Vol.20 No.2

Objectives This study aimed at investigating the role of the proprotein convertase subtilisin/Kexin type 9 (PCSK9)-mediated autophagy on myocardial ischemia/reperfusion injury (MIRI). To determine the relationship between autophagy, apoptosis, fibrosis, and inflammation in the myocardium, to provide experience in preventing and treating the myocardial ischemia/reperfusion (I/R) injury. Methods An AC16 hypoxia-reoxygenation model and a rat myocardial ischemia–reperfusion model were established. The concentrations of cardiac troponin T (cTnT) and creatine kinase-MB (CKMB) in plasma were measured by ELISA. To determine the size of the myocardial infarction, TTC/EB staining was performed. In addition to identifying pathological changes in myocardial tissue, Masson’s trichrome stains and H&E stains were used to identify pathological changes. Echocardiography was employed to detect cardiac function. Western blot analysis was then performed to detect the protein expression of Parkin, Pink1, and markers associated with autophagy (Beclin-1, p62, LC3). Results A significant increase in PCSK9 was observed in the myocardium during H/R. In the cardiac-specific PCSK9 knockdown model, cardiac autophagy was significantly inhibited, whereas cardiac-specific PCSK9 overexpression promoted cardiac autophagy. In vivo studies have demonstrated a significant decrease in cardiac autophagy when the PCSK9 inhibitor was administered. Apoptosis induced by I/R was greatly decreased, and myocardial infarction size and function were both improved by PCSK9 inhibitors. Mechanistically, the PCSK9 inhibitor improved the degree of myocardial fibrosis and inhibited the development of inflammation. Conclusions Our results demonstrated that increased PCSK9 via the parkin/pink1 signaling pathway contributes to I/R and H/R by exaggerating excessive autophagy during reperfusion/reoxygenation. In addition, the PCSK9 inhibitor blocked the development of inflammation and improved Infarct size, myocardial function, and myocardial fibrosis.

Multicomponent assessment and ginsenoside conversions of Panax quinquefolium L. roots before and after steaming by HPLC-MSⁿ

Huang, Xin,Liu, Yan,Zhang, Yong,Li, Shuai-Ping,Yue, Hao,Chen, Chang-Bao,Liu, Shu-Ying The Korean Society of Ginseng 2019 Journal of Ginseng Research Vol.43 No.1

Background: The structural conversions in ginsenosides induced by steaming or heating or acidic condition could improve red ginseng bioactivities significantly. In this paper, the chemical transformations of red American ginseng from fresh Panax quinquefolium L. under steaming were investigated, and the possible mechanisms were discussed. Methods: A method with reversed-phase high-performance liquid chromatography coupled with linear ion trap mass spectrometry ($HPLC-MS^n$)-equipped electrospray ionization ion source was developed for structural analysis and quantitation of ginsenosides in dried and red American ginseng. Results: In total, 59 ginsenosides of protopanaxadiol, protopanaxatriol, oleanane, and ocotillol types were identified in American ginseng before and after steaming process by matching the molecular weight and/or comparing $MS^n$ fragmentation with that of standards and/or known published compounds, and some of them were determined to be disappeared or newly generated under different steaming time and temperature. The specific fragments of each aglycone-type ginsenosides were determined as well as aglycone hydrated and dehydrated ones. The mechanisms were deduced as hydrolysis, hydration, dehydration, and isomerization of neutral and acidic ginsenosides. Furthermore, the relative peak areas of detected compounds were calculated based on peak areas ratio. Conclusion: The multicomponent assessment of American ginseng was conducted by $HPLC-MS^n$. The result is expected to provide possibility for holistic evaluation of the processing procedures of red American ginseng and a scientific basis for the usage of American ginseng in prescription.

Stabilized finite element technique and its application for turbulent flow with high Reynolds number

Huang, Cheng,Yan, Bao,Zhou, Dai,Xu, Jinquan Techno-Press 2011 Wind and Structures, An International Journal (WAS Vol.14 No.5

In this paper, a stabilized large eddy simulation technique is developed to predict turbulent flow with high Reynolds number. Streamline Upwind Petrov-Galerkin (SUPG) stabilized method and three-step technique are both implemented for the finite element formulation of Smagorinsky sub-grid scale (SGS) model. Temporal discretization is performed using three-step technique with viscous term treated implicitly. And the pressure is computed from Poisson equation derived from the incompressible condition. Then two numerical examples of turbulent flow with high Reynolds number are discussed. One is lid driven flow at Re = $10^5$ in a triangular cavity, the other is turbulent flow past a square cylinder at Re = 22000. Results show that the present technique can effectively suppress the instabilities of turbulent flow caused by traditional FEM and well predict the unsteady flow even with coarse mesh.

Baicalein Induces Programmed Cell Death in Candida albicans

Bao Di Dai,Ying Ying Cao,Shan Huang,Yong Gang Xu,Ping Hui Gao,Yan Wang,Yuan Ying Jiang 한국미생물 · 생명공학회 2009 Journal of microbiology and biotechnology Vol.19 No.8