Objective : To explore the role and molecular mechanism of circHIPK3 in the migration and invasion of nasopharyngeal carcinoma cells. Methods : Firstly , we searched public databases and analyzed the expression level of circHIPK3 in nasopharyngeal carcinoma tissues and non⁃cancerous tissues in Gene Expression Database (GEO) . Then , the circHIPK3 siRNA was synthesized and transfected into the NPC cell line 5 ⁃8F. After validate the efficiency of circHIPK3 knockdown , the proliferation , migration and invasion capacities of 5 ⁃8F cells were evaluated by CCK⁃8 , wound healing and transwell assay. Meanwhile , the mRNA and protein expression of downstream genes , including CDH1 , CDH2 , MMP2 , MMP9 and IL⁃6/STAT3 were determined by RT⁃qPCR and Western blot. Finally , transcriptome sequencing technology was used to analyze the significantly differentially expressed genes after the interference of circHIPK3 ⁃siRNA in 5 ⁃8F cells , in order to find out the key genes mediating the oncogenesis role of circHIPK3. Results : circHIPK3 expression significantly increased in nasopharyngeal carcinoma tissues compared with non⁃cancerous tissues ( P < 0. 05 ) . After siRNA⁃specific knockdown of circHIPK3 expression in 5 ⁃8F cells , the proliferation , scratch healing rate , and the number of migrated /invaded cells were significantly reduced (P < 0. 05 , P < 0. 01 , P < 0. 05 respectively) . Meanwhile , the mRNA and protein levels of CDH1 significantly increased (P < 0. 001) , but the CDH2 , MMP2 , MMP9 , IL⁃6/STAT3 mRNA and protein levels were significantly reduced (P < 0. 05) . Further analysis by RNA⁃seq showed that the expression of AL645922. 1 , SCO2 , AL136295. 1 and ISY1 ⁃RAB43 was significantly up⁃regulated( P < 0. 05 ) , and the expression of BIVM⁃ERCC5 , FAM47E⁃ST⁃BD1 , INO80B⁃WBP1 , NAIP , C15orf38⁃AP3S2 , BCL2L2⁃PABPN1 and SMIM11B was significantly down⁃regulated (P < 0. 05) in circHIPK3 ⁃siRNA transfected 5 ⁃8F cells. Conclusion circHIPK3 expression is significantly upregulated in nasopharyngeal carcinoma tissues. circHIPK3 promotes 5 ⁃8F cells proliferation , migration and invasion by regulating the expression of E ⁃Cadherin , N ⁃Cadherin , MMP2 , MMP9 and IL⁃6/STAT3. Genes such as AL645922.1 and BIVM⁃ERCC5 may play a key role in promoting the migration and invasion of nasopharyngeal carcinoma 5 ⁃8F cells mediated by circHIPK3.

Animal survival necessitates adaptive behaviors in volatile environmental contexts. Virtual reality (VR) technology is instrumental to study the neural mechanisms underlying behaviors modulated by environmental context by simulating the real world with maximized control of contextual elements. Yet current VR tools for rodents have limited flexibility and performance (e.g., frame rate) for context-dependent cognitive research. Here, we describe a high-performance VR platform with which to study contextual behaviors immersed in editable virtual contexts. This platform was assembled from modular hardware and custom-written software with flexibility and upgradability. Using this platform, we trained mice to perform context-dependent cognitive tasks with rules ranging from discrimination to delayed-sample-to-match while recording from thousands of hippocampal place cells. By precise manipulations of context elements, we found that the context recognition was intact with partial context elements, but impaired by exchanges of context elements. Collectively, our work establishes a configurable VR platform with which to investigate context-dependent cognition with large-scale neural recording.
Animals ; Mice ; Rodentia ; Virtual Reality ; Cognition ; Recognition, Psychology

Elucidating the active components of traditional Chinese medicine(TCM)is essential for understanding the mechanisms of TCM and promote its rational use as well as TCM-derived drug development.Recent studies have shown that surface plasmon resonance(SPR)technology is promising in this field.In the present study,we propose an SPR-based integrated strategy to screen and analyze the major active components of TCM.We used Radix Paeoniae Alba(RPA)as an example to identify the compounds that can account for its anti-inflammatory mechanism via tumor necrosis factor receptor type 1(TNF-R1).First,RPA extraction was analyzed using an SPR-based screening system,and the potential active in-gredients were collected,enriched,and identified as paeoniflorin and paeonol.Next,the affinity con-stants of paeoniflorin and paeonol were determined as 4.9 and 11.8 μM,respectively.Then,SPR-based competition assays and molecular docking were performed to show that the two compounds could compete with tumor necrosis factor-α(TNF-α)while binding to the subdomain 1 site of TNF-R1.Finally,in biological assays,the two compounds suppressed cytotoxicity and apoptosis induced by TNF-α in the L929 cell line.These findings prove that SPR technology is a useful tool for determining the active in-gredients of TCM at the molecular level and can be used in various aspects of drug development.The SPR-based integrated strategy is reliable and feasible in TCM studies and will shed light on the eluci-dation of the pharmacological mechanism of TCM and facilitate its modernization.

Diabetic nephropathy (DN) is currently the most common complication of diabetes. It is considered to be one of the leading causes of end-stage renal disease (ESRD) and affects many diabetic patients. The pathogenesis of DN is extremely complex and has not yet been clarified; however, in recent years, increasing evidence has shown the important role of innate immunity in DN pathogenesis. Pattern recognition receptors (PRRs) are important components of the innate immune system and have a significant impact on the occurrence and development of DN. In this review, we classify PRRs into secretory, endocytic, and signal transduction PRRs according to the relationship between the PRRs and subcellular compartments. PRRs can recognize related pathogen-associated molecular patterns (PAMPs) and danger-associated molecular patterns (DAMPs), thus triggering a series of inflammatory responses, promoting renal fibrosis, and finally causing renal impairment. In this review, we describe the proposed role of each type of PRRs in the development and progression of DN.
Alarmins/physiology* ; C-Reactive Protein/physiology* ; Diabetic Nephropathies/etiology* ; Endocytosis ; Humans ; Immunity, Innate ; Mannose-Binding Lectin/physiology* ; Pathogen-Associated Molecular Pattern Molecules ; Receptors, Pattern Recognition/physiology* ; Serum Amyloid P-Component/physiology* ; Signal Transduction

Some unhealthy life habits, such as long-term smoking, heavy drinking, sexual overstrain and frequent stay-up could induce the Yin deficiency symptoms of zygomatic red and dysphoria. Stems of Dendrobii officinalis flos (DOF) showed the efficacy of nourishing Yin. In this study, the hyperthyroidism Yin deficiency model was set up to study the yin nourishing effect and action mechanism of DOF, in order to provide the pharmacological basis for developing DOF resources and decreasing resource wastes. ICR mice were divided into five groups: the normal control group, the model control group, the positive control group and DOF extract groups (6.4 g · kg(-1)). Except for the normal group, the other groups were administrated with thyroxine for 30 d to set up the hyperthyroidism yin deficiency model. At the same time, the other groups were administrated with the corresponding drugs for 30 d. After administration for 4 weeks, the signs (facial temperature, pain domain, heart rate and autonomic activity) in mice were measured, and the facial and ear micro-circulation blood flow were detected by laser Doppler technology. After the last administration, all mice were fasted for 12 hours, blood were collected from their orbits, and serum were separated to detect AST, ALT, TG and TP by the automatic biochemistry analyzer and test T3, T4 and TSH levels by ELISA. (1) Compared with the normal control group, the model control group showed significant increases in facial and ear micro-circulation blood flow, facial temperature and heart rate (P < 0.05, P < 0.01), serum AST, ALT (P < 0.01), T3 level (P < 0.05), TSH level (P < 0.05) and notable deceases in pain domain (P < 0.01), TG level (P < 0.01). (2) Compared with the model control group, extracts from DOF (6 g · kg(-1)) could notably reduce facial and ear micro-circulation blood flow, facial temperature and heart rate (P < 0.05, P < 0.01) and AST (P < 0.05) and enhance pain domain (P < 0.01) and TG (P < 0.01). Extracts from DOF (4 g · kg(-1)) could remarkably reduce AST and ALT levels (P < 0.01, 0.05). Extracts from DOF (6 g · kg(-1) 4 g · kg(-1)) could significantly reduce T3 and increase serum TSH level (P < 0.05). DOF could improve Yin deficiency symptoms of zygomatic red and dysphoria in mice as well as liver function injury caused by overactive thyroid axis. According to its action mechanism, DOF may show yin nourishing and hepatic protective effects by impacting thyroxin substance metabolism, improving micro-circulation and reducing heart rate.
Animals ; Dendrobium ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Flowers ; chemistry ; Humans ; Hyperthyroidism ; drug therapy ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Phytotherapy ; Thyroxine ; metabolism ; Yin Deficiency ; drug therapy ; metabolism

Objective To develop a method for determination of 13 kinds of bile acid in healthy people and patients with pri-mary liver cancer by HPLC-MS.Methods Acetonitrile was used to precipitate protein in plasma samples .The separation was per-formed on a SHISEIDO MG C18 reverse phase column (3.0 mm ×100 mm,3μm).The mobile phase were acetonitrile and water (con-taining 0.1%formic acid and 0.5%ammonia), gradient elution.The flow rate was 0.4 ml/min, the temperature of column was 25℃and the injection volume was 5 μl.Electrospray ion source (ESI) and selected ion mode (SIM) were applied for quantitative analysis under negative ion mode .Results The calibration curves of 13 kinds of bile acids were linear in the range of 3.3-4 400 ng/ml, intra-day and inter-day precision results were all less than 7%, the average recovery were 82.67%-109.75%.The result showed that the content of conjugated bile acid increased in the plasma of patients with primary liver cancer compared with the healthy control group , especially in TUDCA, TCDCA, TCA, GCA and GCDCA.Conclusion The LC -MS method was sensitive and selective which could be suitable for determination of bile acid in healthy people and patients with primary liver cancer .The study laid a foundation for the me-tabolism of bile acid in vivo and the clinical diagnosis and treatment of patients with hepatocellular carcinoma .

Genetic studies are traditionally based on single-gene analysis. The use of these analyses can pose tremendous challenges for elucidating complicated genetic interplays involved in complex human diseases. Modern pathway-based analysis provides a technique, which allows a comprehen-sive understanding of the molecular mechanisms underlying complex diseases. Extensive studies uti-lizing the methods and applications for pathway-based analysis have significantly advanced our capacity to explore large-scale omics data, which has rapidly accumulated in biomedical fields. This article is a comprehensive review of the pathway-based analysis methods––the powerful methods with the potential to uncover the biological depths of the complex diseases. The general concepts and procedures for the pathway-based analysis methods are introduced and then, a comprehensive review of the major approaches for this analysis is presented. In addition, a list of available path-way-based analysis software and databases is provided. Finally, future directions and challenges for the methodological development and applications of pathway-based analysis techniques are dis-cussed. This review will provide a useful guide to dissect complex diseases.

Objective:This study aimed to clarify the correlation of SPRY4-IT1 expression with the clinicopathological character-istics and prognosis of patients with esophageal squamous cell carcinoma (ESCC), as well as the role of SPRY4-IT1 in promoting ES-CC cell growth. Methods:Quantitative real-time polymerase chain reaction for SPRY4-IT1 expression was performed on 50 paired can-cerous and adjacent non-cancerous esophageal specimens. Small interfering RNA was used to suppress SPRY4-IT1 expression to fur-ther explore its role in tumor progression. Cell viability was tested in vitro by MTT assay (OD=490 nm), and cell apoptosis and cell cy-cle were investigated by flow cytometry. Results:We found markedly elevated SPRY4-IT1 expression in cancerous tissues compared with adjacent non-cancerous tissues (90%, P<0.01). Relative SPRY4-IT1 expression levels were correlated with some clinicopathologi-cal characteristics, such as tumor size (χ2=5.333, P=0.021), elevated TNM (2009) stage classi fi cation (χ2=5.556, P=0.018), and de-creased overall survival rates (χ2=5.296, P=0.021). SPRY4-IT1 expression level was not correlated with patient age, gender, smoking status, or alcohol consumption (all P>0.05). Further experiments showed that SPRY4-IT1 expression levels were significantly higher in three ESCC cell lines than in the normal human esophageal epithelial cell line Het-1A. In vitro assays of the ESCC cell line KYSE30 demonstrated that knockdown of SPRY4-IT1 expression by small interfering RNA reduced cell growth, mediated cell cycle arrest at the G0-G1 phase, and promoted cell apoptosis (all P<0.01). Conclusion:SPRY4-IT1 was overexpressed in ESCC tissues and ESCC cell lines and promoted the growth of ESCC cells. The dysregulated expression of long non-coding RNA SPRY4-IT1 may play an important role in the process of ESCC development and may be developed as a useful biomarker for the diagnosis and prognosis of ESCC.

OBJECTIVETo observe the effect of vascular endothelial growth factor (VEGF) on bone marrow-derived mesenchymal stem cell (MSC) proliferation and explore the signaling mechanism involved.

METHODSMSC culture was performed following the classical whole bone marrow adhering method. The characteristics of MSC were identified by induction of multi-lineage differentiation and flow cytometry for surface marker analysis (CD34, CD45, CD29, and CD90). Following the addition of 50 nmol/L wortmannin, 50 µmol/L PD98059, 30 µmol/L SB203580, 10 µmol/L H89, 20 µmol/L Y27632, 1 µmol/L rapamycin, 10 µmol/L straurosporine, 6 nmol/L Go6976, or 50 µmol/L Pseudo Z inhibitors in the cell culture, the MSC were treated with 20 ng/ml VEGF and the changes of the cell proliferation rate was measured with MTT assay.

RESULTSCultured MSC were capable of multi-linage differentiation and did not express VEGF-R, CD29 or CD90. Treatment with 20 ng/ml VEGF obviously promoted MSC proliferation, and this effect was inhibited partially by p38 mitogen-activated protein kinase (MAPK) inhibitor rapamycin, PD98059, SB203580, Go6976, and straurosporine.

CONCLUSIONSVEGF promotes MSC proliferation in close relation to the AKT-PKC pathway, in which PKC signal pathway may play the central role.

Animals ; Bone Marrow Cells ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Female ; Male ; Mesenchymal Stromal Cells ; cytology ; Protein Kinase C ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVETo explore the effects of 1,2-dichloroethane (1,2-DCE) on the behavior and the brain neurotransmitter levels in mice.

METHODSThirty mice were randomly divided into four groups, which were control group and groups of low, middle and high exposure (225, 450 and 900 mg/m3) to 1,2-DCE for 10 days (3.5 h a day) by inhalation. After the last exposure, the open field test was performed immediately. After exposure all mice were killed and the brain tissues were taken up rapidly. The levels of aspartate (Asp), glutamate (Glu) and gamma-aminobutyric acid (GABA) in the brain were detected by high performance liquid chromatography (HPLC).

RESULTSLevels of Asp and Glu in all exposure groups increased with doses. As compared to the control group, levels of Glu in all exposure groups increased significantly (P < 0.05). Levels of GABA in the low exposure group were significantly lower than those in control group, but those in the high exposure group were significantly higher than those in control group. The results of the open field test showed that effect of low exposure to 1,2-DCE on the behavior was stimulant, but the high exposure to 1,2-DCE inhibited behavior of exploration, excitement and sport.

CONCLUSIONSSubacute exposure to 1,2-DCE could result in the change of amino acid neurotransmitter content and ratio in the brain, thereby change the behavior of mice appeared, which might be the mechanism of neurotoxicity caused by 1,2-DCE in part.

Animals ; Aspartic Acid ; analysis ; Behavior, Animal ; drug effects ; Brain ; metabolism ; Ethylene Dichlorides ; toxicity ; Female ; Glutamic Acid ; analysis ; Mice ; Mice, Inbred Strains ; Neurotransmitter Agents ; metabolism ; gamma-Aminobutyric Acid ; analysis