Objective To identify mutations of CACNA1A gene in a family with hemiplegic migraine.Methods Total genomic DNA was extracted from a family with 3 affected members and 1 000 healthy controls.The proband and his patient sister were subjected to exome sequencing.Ten family members including 3 patients were subjected to linkage analysis.The coding exons of the CACNA1A gene were amplified and sequenced in affected and normal individuals. Bioinformatics analysis were performed.Results A novel CACNA1A mutation was identified in the 3 patients.The nonsense mutation of A to G was detected at nucleotide 1168 ( c.1168A >G) which converted the Asn codon ( AAT) to Asp (GAT) in exon 8.Conclusion The mutation(N390D) detected in the present study is considered to result in the Chinese Hemiplegic migraine family.

OBJECTIVE:To investigate the characteristics and causes of cardiac ADR induced by quinolones,and to provide reference for clinical treatment. METHODS:Three thousaud two hundred and eighty-eighe 8 patients receiving common quinolones were selected from clinical departments in Zhengzhou Central Hospital of Zhengzhou University during Mar. 2012-Mar. 2016. Retro-spective analysis was conducted in terms of patients’age and gender,clinical departments,main clinical manifestations,route of administration,underlying disease,drug combination. The reasons for cardiac ADR were analyzed. RESULTS:Among 3 288 pa-tients,there were 34 patients(1.03%)with cardiac ADR. Among them,the incidence of cardiac ADR in patients over 50 years old was as high as 76.47%;patients with cardiac ADR were mainly in the respiration department,gastroenterology department and urology department,accounting for 76.47%;most of patients were from gastroenterology department(29.41%). In cardiac ADR, the main clinical manifestations were QTc interval prolongation torsadesde poiutes(TdP)and TdP,accounting for 58.82%. Among them,patients with QTc interval prolongation TdP occupied the highest proportion,there was no statistical significance compared to TdP(P>0.05);there was statistical significance in the difference with other clinical manifestations(P<0.05). Among common-ly used quinolones,levofloxacin(32.35%)and ciprofloxacin(41.18%)caused large proportion of cardiac ADR,there was statisti-cal significance compared to norfloxacin,moxifloxacin and other quinolones(P<0.05). The proportion of cardiac ADR induced by intravenous dripping(91.18%)was much higher than oral administration(8.82%),with statistical significance(P<0.05). Among patients with cardiac ADR,the patients with underlying disease (94.12%) and drug combination (91.18%) occupied the higher proportion,there was statistical significance compared to the patients without underlying disease and drug combination(P<0.01). Among drug combination,the patients receiving amiodarone (29.41%) and salbutamol (20.59%) occupied the large proportion, there was statistical significance compared to other types of drug combination(P<0.05). CONCLUSIONS:Cardiac ADR induced by quinolones in our hospital mostly occurs in respiration department,gastroenterology department and urology department,and mainly manifests as QTc interval prolongation TdP and TdP. The incidence of cardiac ADR may be greatly increased in elderly pa-tients,patients with underlying diseases,and drug combination as well as intravenous infusion. Therefore,clinicians should select suitable quinolones,and make reasonable individualized dosage regimen.

Objective To investigate the relationship between familial hemiplegic migraine (FHM ) with the mutation of CACNA1A gene .Methods Total genomic DNA of a family affected members and 1 000 normal controls was extracted for conduc‐ting the CACNA1A gene sequencing research and the bioinformatics analysis .Results The novel mutation site c .1168A>G of ex‐on located in CACNA1A gene led to Asn to be replaced with Asp (N390D) .Conclusion The mutation(N390D) of CACNA1A gene is a newly found novel pathogenic mutation lead to familial hemiplegic migraine .

AIMTo investigate the effects of prostaglandin E 1 on apoptosis induced by hypoxia/reoxygenation in cultured neonatal rat cardiomyocy tes. METHODSThe models of hypoxia/reoxygenation were made with t he first generation of cultured cardiomyocytes. Hypoxia/reoxygenation apoptosis in cultured neonatal rat cardiomyocytes was studied by agarose gel electrophores is and Tdt-mediated dUTP nick end labeling(TUNEL). Bcl-2 and bax mRNA were det ected by in situ hybridization. RESULTSThe results of DNA electr ophoresis in the H/R group showed the typical DNA ladder. And the DNA ladder decreased gradually corresponding to the increased dose of PGE 1. The TUNEL staining showed that the total number of apo ptotic cells in the H/R group was much biger than that in PGE 1(0 127 ?mol?L -1 ) group. The results of in situ hybridization showed that the conten t of bcl-2 mRNA in H/R group was lower than control. And the content of bax mRN A showed a reverse result as bcl-2 mRNA. Compared with H/R group, the content o f bcl-2 mRNA was significantly higher after treatment with PGE 1(0 014 ?mol ?L -1 , 0 042 ?mol?L -1 , 0 127 ?mol?L -1 ). But the content of bax mRNA in PGE 1(0 014 ?mol?L -1 , 0 042 ?mol?L -1 , 0 127 ?mol?L -1 )groups was significantly lower than H/R group. CONCLUSI ONH/R injury can induce cardiomyocyte apoptosis. PGE 1 has obvious an ti-apoptotic effects on cardiomyocyte and the mechanisms are possibly by inhibi ting the expression of bax and increasing the expression of bcl-2.hein creaseddoseofPGE1 .TheTUNELstainingshowedthatthetotalnumberofapoptoticcellsintheH

Objective To study the anti-inflammatory effects of idazoxan (IDA) on endotoxin lipopolysaccharide (LPS) challenged mice in vivo and activated macrophages in vitro,and explore its potential molecular mechanisms.Methods To do the experiments in vivo,30 adult male C57BL/6 mice were randomly divided into control group,model group,and low,medium and high doses IDA groups (IDA-L,IDA-M,and IDA-H groups),n =6 in each group.The inflammatory model was reproduced by intraperitoneal injection of LPS 10 mg/kg,and the control group was injected with the same amount of normal saline.The IDA groups received LPS (10 mg/kg) and IDA 0.3,1.0 and 3.0 mg/kg,respectively.The blood samples of mice in each group were collected at 6 hours after the reproduction of the model.For the in vitro experiments,primary peritoneal macrophages were collected from 20 adult male C57BL/6 mouse cells and they were divided into control group,LPS group (10 mg/L) and LPS+IDA-L,IDA-M,IDA-H groups (10 mg/L LPS + 5,25,100 μmol/L IDA,respectively).Cell culture supernatants were collected at 24 hours after the reproduction of the model.Detection methods:enzyme linked immunosorbent assay (ELISA) was used to determine the levels of serum tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),monocyte chemotactic protein-1 (MCP-1) and nitric oxide (NO).Western Blot was used to determine the effect of IDA on the expression levels of nuclear factor-κB (NF-κB) in macrophages.Results ① For the in vivo experiment,the serum levels of TNF-α and IL-6 were significantly elevated in the model group as compared with those in the control group [TNF-o (ng/L):403.96 ± 40.98 vs.17.50 ± 8.68;IL-6 (ng/L):61 400.31 ± 7 826.61 vs.2 436.30 ± 448.89;both P ＜ 0.01].IDA treatment could inhibit the elevation of inflammatory cytokines in a dose-dependent manner,with the most significant decrease in LPS+IDA-H group [TNF-α (ng/L):170.09 ± 28.53 vs.403.96 ± 40.98,IL-6 (ng/L):16 570.81 ± 1 083.65 vs.61 400.31± 7 826.61;both P ＜ 0.01].② For the in vitro experiment,the levels of TNF-α,IL-6,MCP-1,and NO secreted by LPS-stimulated macrophages were distinctly higher in the LPS group than those in the control group [TNF-α (ng/L):7 259.14 ± 320.70 vs.28.50±27.08,IL-6 (ng/L):14809.60±5852.73 vs.1 113.47±465.53,MCP-1 (ng/L):20847.37± 1 788.33 vs.447.37± 395.69,NO (μmol/L):1 900.00 ± 144.31 vs.603.03 ± 102.18;all P ＜ 0.01].However,IDA intervention could lower the secretion of TNF-α,IL-6,MCP-1 and NO in a dose-dependent manner,with the most notable decrease in the LPS+IDA-H group [TNF-α (ng/L):784.40±281.90 vs.7259.14±320.70,IL-6 (ng/L):1 802.96± 1 534.18 vs.14 809.60± 5 852.73,MCP-1 (ng/L):2005.26± 1 534.28 vs.20847.37 ± 1 788.33,NO (μ mol/L):654.54± 150.21 vs.1 900.00 ± 144.31;all P ＜ 0.05].In addition,IDA at the concentration of 100 μmol/L could promote the translocation of NF-κBp65 in macrophages into the nucleus 15 minutes early and lead to increased NF-κBp65 expression (gray value:18.70 ± 2.29 vs.1.09 ± 0.36,P ＜ 0.05),hut significantly reduce the expression levels of NF-κBp50 in the nucleus at 45 minutes after treatment (gray value:1.99 ± 0.14 vs.2.94 ± 0.54,P ＜ 0.05).Conclusions IDA could significantly reduce inflammation of mice challenged with LPS and inhibit inflammatory cytokines and mediators secreted by macrophage in a dose-dependent manner.High concentration of IDA (100 μmol/L) exhibited the greatest anti-inflammatory effects.The anti-inflammatory effect of IDA may be worked through NF-κB signaling pathway.

Objective To compare the effects of telmisartan and (or) amlodipine on the reversal left ventricular re-modeling in two-kidney one clip hypertensive rats. Methods A total of 50 healthy male SD rats were randomly divided into 5 groups (n=10):two-kidney one clip renal hypertensive (2KIC) model group, sham group, telmisartan (10 mg/kg) group, am-lodipine (2.5 mg/kg) group and telmisartan (10 mg/kg)+amlodipine(2.5 mg/kg) group. The model of two-kidney one clip re-nal hypertensive rats was established. The tail arterial blood pressure was detected once a week. After 20 weeks, rats were sacrificed and specimens were collected. The left ventricular mass index (LVMI) was assessed. The myocardial ultrastructur-al changes were observed by electron microscope. Values of plasma renin activity (PRA), angiotensionⅡ(AngⅡ) and atrial natriuretic peptide (ANP) were measured by enzyme linked immunosorbent assay (ELISA).Results Compared with sham group, the levels of systolic blood pressure (SBP), LVMI, PRA, AngⅡand ANP were significantly higher in 2KIC group (P＜0.01). Compared with 2KIC group, values of SBP, LVMI, PRA and ANP were significantly lower in telmisartan group and am-lodipine group (P＜0.01), but the value of AngⅡwas significantly higher (P＜0.01). The levels of SBP, LVMI, AngⅡand ANP were significantly lower in combined medication group than those of single drug medication group (P＜0.01). There was no significant difference in the plasma PRA level between those groups (P>0.05). Results of myocardial electron microsco-py showed that the left ventricular remodeling was significantly improved in combined treatment group. Conclusion Telmisartan and amlodipine can effectively improve the left ventricular remodeling induced by hypertension. There was more effective therapy using both medications together.

When the size of a neurosphere cultured in vitro reaches a certain critical value, a necrotic core will appear inside the neurosphere because of the limitation of oxygen or other nutrients transport from medium to the cells in the neurasphere. Large necrotic core will greatly reduce the expansion of NSCs. The cellular automaton (CA) model is applied in this article to model the growth of NSCs in sphere state. The appearance and enlargement of the necrotic core in a neurosphere is calculated by coupling the CA model with the nutrient diffusion analysis in bioreactors. The calculation results indicate that the culture conditions, such as seeding density, the concentration of nutrients in medium and the mass transfer coefficient between a neurosphere and medium, have some effects on the appearance of the necrotic core. However, the necrotic core mainly depends on the inner diffusion. It will certainly appear if the size of the neurosphere is large enough even the outside mass transfer is in a good condition in bioreactors. Additionally, the appearance of the necrotic core resulting from the shortage of oxygen is earlier than that caused by the limitation of glucose. And the growth of the necrotic core is very fast after its appearance, and the whole neurosphere may become necrotic. The model developed with cellular automaton and mass transfer is a good qualitative representation of NSCs growth in bioreactors.
Bioreactors ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Computer Simulation ; Models, Biological ; Neurons ; cytology ; Spheroids, Cellular ; Stem Cells ; cytology ; Tissue Engineering ; methods

Objective To find out whether conversation analysis helps to differentiate psychogenic nonepileptic seizure (PNES) from epileptic seizure in Chinese patients.Methods Twelve unselected patients from Peking Union Medical College Hospital during 2014 to 2016 with diagnostic uncertainty were included.Interactions following standard protocol were carried out.A linguist blinded to all medical data and a neurologist studied videos and transcripts of the interactions.Using a diagnostic scoring aid which includes 17 conversation features summarized from previous researches, they attempted to predict the medical diagnosis of those patients independently.Results Accurate diagnosis was predicted in 10/12 patients by both raters.Average scores of patients with epileptic seizures were 8.00 (linguist) and 6.75 (neurologist), while average scores of paitents with PNES were-5.75 (linguist) and-7.88 (neurologist).Both raters agreed on most individual items (81.86%, 167/204).To demonstrate different features between these two groups, a case comparison was made between one patient with frontal lobe epilepsy and one patient with PNES.Conclusion In Chinese patients, conversation analysis can help differentiate between epileptic seizure and PNES.

Breast cancer is the most common cancer in women and one of the deadliest cancers worldwide.According to the distribution of tumor tissue,breast cancer can be divided into invasive and non-invasive forms.The cancer cells in invasive breast cancer pass through the breast and through the immune system or systemic circulation to different parts of the body,forming metastatic breast cancer.Drug resistance and distant metastasis are the main causes of death from breast cancer.Research on breast cancer has attracted extensive attention from researchers.In vitro construction of tumor models by tissue engineering methods is a common tool for studying cancer mechanisms and anticancer drug screening.The tumor microenvironment consists of cancer cells and various types of stromal cells,including fibroblasts,endothelial cells,mesenchymal cells,and immune cells embedded in the extracellular matrix.The extracellular matrix contains fibrin proteins(such as types Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅵ,and Ⅹcollagen and elastin)and glycoproteins(such as proteoglycan,laminin,and fibronectin),which are involved in cell signaling and binding of growth factors.The current traditional two-dimensional(2D)tumor models are limited by the growth environment and often cannot accurately reproduce the heterogeneity and complexity of tumor tissues in vivo.Therefore,in recent years,research on three-dimensional(3D)tumor models has gradually increased,especially 3D bioprinting models with high precision and repeatability.Compared with a 2D model,the 3D environment can better simulate the complex extracellular matrix components and structures in the tumor microenvironment.Three-dimensional models are often used as a bridge between 2D cellular level experiments and animal experiments.Acellular matrix,gelatin,sodium alginate,and other natural materials are widely used in the construction of tumor models because of their excellent biocompatibility and non-immune rejection.Here,we review various natural scaffold materials and construction methods involved in 3D tissue-engineered tumor models,as a reference for research in the field of breast cancer.

Objective:To investigate the chemical constituents in the rattan of Rubia argyi L.. Methods:The air-dried rattan of Rubia argyi L. was powdered and extracted three times by 75% ethanol with refluxing. After removing the solvent under the reduced pressure,the crude extract was dissolved in water,and then filtrated and extracted by petroleum ether and ethyl acetate to obtain crude extract after removing petroleum ether and ethyl acetate. The compounds were isolated and purified by silica gel column chromatogra-phy,reversed-phase silica gel column chromatography and Sephadex LH-20 gel column chromatography,and then identified based on physicochemical properties and spectral analysis(1 H-NMR and 13C-NMR). Results:Totally 13 compounds were isolated from the rat-tan of Rubia argyi L.,and characterized as secoisolariciresinol(1),xanthopurpurin(2),daucosterol(3),dehydroabietic acid(4), 2-hydroxy-1-methoxy-anthraquinone(5),β-sitosterol(6),lirioresinol A(7),2-hydroxy-7-methyl-9,10-anthraquinone(8),strych-novoline (9), ciwujiatone (10), 3,4-divanillyltetrahydrofuran (11), 2-(4-hydroxypheny) -6-(3-methoxy-4-hydroxyphenyl)-3,7-dioxabicyclo[3,3,0]octane (12), and (6S,9R)-vomifoliol (13).Conclusion: The compounds 1-13 are isolated from the rattan of Rubia argyi L. for the first time and the compounds 1,2,4,5 and 7-13 are first isolated from Rubia L..