1.Altered expression and clinical significance of plasma circulating microRNA-23b in patients with myasthnia gravis
Xueling ZHOU ; Jianjian WANG ; Huixue ZHANG ; Lihua WANG
Chinese Journal of Neurology 2017;50(3):213-217
Objective To evaluate the expression and clinical significance of plasma circulating microRNA-23b in patients with myasthenia gravis (MG).Methods Plasma samples from 32 MG patients and 32 healthy people were collected in the Second Affiliated Hospital of Harbin Medical University from October 2015 to August 2016. Expression of microRNA-23b was detected by quantitative reverse transcription polymerase chain reaction .The relationship among microRNA-23b expression level, clinical features and quantitative myasthenia gravis score ( QMG ) was analyzed. Results The expression of circulating microRNA-23b in plasma was significantly higher in MG patients (4.587 (3.654, 5.423)) compared with control group (2.889 (1.968, 4.027), Z=-4.169, P<0.01), meanwhile it was positively related to QMG (r=0.661,P<0.01).But the expression of circulating microRNA-23b had no statistically significant difference in MG subgroups .Conclusion Plasma circulating microRNA-23b is highly expressed in MG and has positive correlation with MG severity .
2.Inhibitory effects of bone sialoprotein monoclonal antibody on bone-seeking breast cancer cells adhering to bone matrix
Yingjing GONG ; Jie WANG ; Huixue SONG ; Bing XIA
Chinese Journal of Cancer Biotherapy 2006;0(06):-
Objective: To determine the inhibitory effects of anti-bone sialoprotein(BSP) antibody on human breast cancer cells adhering to bone matrix in vitro.Methods: Expression of BSP in 3 different cancer cell lines(bone seeking breast cancer cell MDA-MB-231-BO,lung adenocarcinoma cell SPCA-1 and colon carcinoma cell LOVO)was detected immunohistochemically.Cells adhering test was carried out to investigate the adhering of the 3 different cancer cell lines to mouse bone matrix in vitro and the inhibitory effect of anti-BSP antibody on MDA-MB-231-BO cells adhering to mouse bone matrix;Enzyme-linked immunosorbent assay was carried out for quantitative detection of TGF-?.Results: BSP immunostaining was positive in MDA-MB-231-BO cells and negative in SPCA-1 and Lovo cells.The number of MDA-MB-231-F10 cells adhering to mouse bone matrix was significantly more than SPCA-1 or Lovo cells(P
4.Dura mater spinalis integrity may influence cytokine levels in the cerebrospinal fluid
Wanshan BAI ; Xinwei WANG ; Wen YUAN ; Zhanchao WANG ; Lei LIANG ; Huixue WANG
Chinese Journal of Tissue Engineering Research 2013;(33):6001-6004
BACKGROUND:Pathophysiological mechanisms after spinal cord injury are very complex, so there is no compressive and in-depth understanding on it.
OBJECTIVE:To study the effect of dura mater spinalis integrity on cytokine levels in the cerebrospinal fluid of animal models of spinal cord injury.
METHODS:The white rabbit models of spinal cord injury were established using clamp compression method, and then the models were randomly divided into four groups:no dura mater spinalis defect group, dura mater spinalis defect group, dura mater spinalis defect composite with membrane repairing group and dura mater spinalis defect composite with autologous fascia repair group. Enzyme-linked immunosorbent assay was
performed to detect the changes of levels of cytokines (interleukin-6, interleukin-10 and tumor necrosis factorα) in the cerebrospinal fluid at 30 minutes, 1, 3, 6, 12 and 36 hours after surgery.
RESULTS AND CONCLUSION:The levels of interleukin-6, interleukin-10 and tumor necrosis factorαin the
cerebrospinal fluid of the dura mater spinalis defect group, dura mater spinalis defect composite with membrane repairing group and dura mater spinalis defect composite with autologous fascia repair group were significantly lower than those of the no dura mater spinalis defect group at 6 hours after surgery (P<0.05). There were no significant differences in the levels of interleukin-6, interleukin-10 and tumor necrosis factorαat other time points between groups (P>0.05). The results indicate that maintaining the integrity of dura mater spinalis of the spinal cord injury model can affect the levels of interleukin-6, interleukin-10 and tumor necrosis factorαin the
cerebrospinal fluid, thus inhibiting the inflammatory response.
5.Altered expression of microRNA-27a-3p in the thymus tissue of patients with myasthenia gravis
Qian LI ; Lihua WANG ; Jianjian WANG ; Huixue ZHANG ; Xue SHAN ; Xiaotong KONG
Chinese Journal of Neurology 2016;49(4):302-306
Objective To investigate the expression level of thymus microRNA-27a-3p in patients with myasthenia gravis (MG) and to explore the pathogenesis of MG.Methods Thymus tissue samples from 36 cases were collected from December 2014 to February 2015 in the Second Affiliated Hospital of Harbin Medical University.Nineteen thymus tissue samples of MG group were collected from department of chest surgery,17 thymus tissue samples of control group were collected from department of chest surgery or congenital heart disease patients from department of cardiac surgery.The expression of microRNA-27a-3p in the thymus from 36 patients was determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR),using U6 as housekeeping control.The Wilcoxon Rank-Sum test was used to analyze the relative expression of microRNA-27a-3p of the two groups.Spearman rank correlation was used to determine the correlation coefficient between microRNA-27a-3p and Quantitative Myasthenia Gravis Score (QMGS).Results (1) The expression level of microRNA-27a-3p in thymus was significantly higher in MG group (0.195(0.049,0.714)) compared with control group (0.045(0.004,0.088),Z =-2.646,P =0.008).(2) Nineteen MG patients were included in the study,out of which 7 were ocular myasthenia gravis (OMG) patients and 12 were generalized myasthenia gravis (GMG) patients.The expression of microRNA-27a-3p in GMG patients (0.493 (0.157,1.123)) was significantly higher than that in OMG patients (0.035 (0.008,0.103),Z =-2.620,P =0.009).(3) There was a positive correlation between the expression of microRNA-27a-3p and QMGS (r =0.576,P =0.010).Conclusions The expression of microRNA-27a-3p in thymus is significantly up-regulated in the patients with MG.MicroRNA-27a-3p may be associated with MG severity and significantly elevated in GMG patients compared with OMG patients.
6.Correlation between single nucleotide polymorphisms of multi-loci and multi-genes and myasthenia gravis
Xiaotong KONG ; Lihua WANG ; Jianjian WANG ; Xue SHAN ; Qian LI ; Huixue ZHANG
Chinese Journal of Neurology 2016;49(4):307-311
Objective To investigate the association between myasthenia gravis (MG) and single nucleotide polymorphisms (SNPs) of PTPN22 + 1858C/T,CTLA-4 (+ 49A/G;-1772C/T;-1661A/G),KRAS(rs9226),BCL2(rs4987855) and IGF-1R(rs34804698) genes.Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was adopted to detect the gene types of SNPs in 76 MG patients who were enrolled in the Second Affiliated Hospital of Harbin Medical University from July 2011 to June 2015 and 59 healthy blood donors.Results In MG patients,the frequences of CTLA-4 +49A/G(rs231775) (57.9%) and-1772C/T (rs733618) (43.4%) were higher than that in the healthy controls (22.1%) (x2 =35.252,P =0.000; x2 =4.098,P =0.043).The frequence of CTLA-4 +49A/G in MG patients combined with thymoma (25.6%) was higher than other subgroups (thymic hyperplasia group:13.8%; normal thymus group:18.4%)(x2 =7.564,P=0.006; x2 =7.155,P=0.007).Meanwhile,the frequence of the C-1772 allele was higher in thymoma group (19.7%) compared with other two groups (thymic hyperplasia group:9.86% ; normal thymus group:13.8%) (x2 =5.331,P =0.021 ;x2 =4.411,P =0.036).However,the other SNPs were not associated with the risk of MG.Conclusion There are associations of MG with CTLA-4 + 49A/G and-1772C/T SNPs,but not with PTPN,KRAS,BC12 and IGF-1R SNPs.
7.Posterior short-segment pedicle screw fixation at the injured level for thoracolumbar spine fractures
Yunrong ZHU ; Xiaojian YE ; Jiangmin YU ; Yuquan JIANG ; Huixue WANG ; Chunquan FAN ; Hailong HE ; Guohua XU
Chinese Journal of Trauma 2010;26(3):221-224
Objective To discuss indications,operation method and clinical outcome of posterior short-segment pedicle fixation at the injured level for treatment of thoracolumbar spine fractures.Methods A total of 38 patients with thoracolumbar spine fractures were equally randomized to Group A(treated with classic short-segment pedicle screw fixation)and Group B(treated with short-segment pedicle screw fixation at the injured level)based on fixation methods(19 patients per group).Preoperative and postoperative JOA score,segmental lordosis(Cobb' s angle),R value(anterior fractured vertebral body height/mean normal vertebral body height×100%),VSA score and internal fixation condition were assessed and compared clinically.Results All patients were followed up for 6-37 months(mean 20.5 months),which showed no statistical difference upon Frankel scores of two operation modes,while the segmental lordosis,VAS score and R value in Group B were than those in Group A.There occurred nuts loosening in one patient and screw bending in one in Group A.There was no implant breakage,loosening or emersion in Group B.Conclusion Posterior short-segmental fixation at the injured level is an adequate and effective procedure for compression fractures and mild to moderate burst fractures of the thoracolumbar spine.
8.Studies on pharmacokinetics of nitidine chloride in rabbits.
Huagang LIU ; Dongmei YE ; Huixue HUANG ; Xibin WANG ; Danni LI
China Journal of Chinese Materia Medica 2009;34(11):1406-1409
OBJECTIVEA high performance liquid chromatography (HPLC) method was developed to determine the concentration of nitidine chloride in plasma and successfully applied to study pharmacokinetics after i.v. administration in rabbits.
METHODTwelve rabbits, randomized into 2 groups , were given i.v. at the dose of 4, 6 mg x kg(-1) respectively. Chloramphenicol was used as an internal standard. Nitidine chloride was extracted from plasma with ion pair reagent, and was determined by HPLC.
RESULTThe calibration curves of nitidine chloride was linear in the range of 0.03-2.04 mg x L(-1). Its recoveries were more than 95%, intra-day and inter-day precisions were lower than 6%. The concentration-time curve of nitidine chloride in rabbits after i.v. of 4 and 6 mg x kg(-1) were shown to fit a two-compartment model, the main pharmacokinetic parameters showed no significant difference between the low and high dosage, and the AUC values are directly relative to doses. T1/2alpha were (5.46 +/- 0.89), (4.76 +/- 0.33) min respectively, T1/2beta were (263.33 +/- 16.4), (274.71 +/- 16.52) min respectively, AUC were (46.56 +/- 1.80), (69.19 +/- 2.30) microg x min(-1) x mL(-1) respectively.
CONCLUSIONIt is first time to establish the HPLC method to determine the concentration of nitidine chloride in rabbits plasma. The method is sensitive, accurate and reproducible. It is first time to study the pharmacokinetic characters of nitidine chloride in rabbits after i.v. administration, the elimination of nitidine chloride is linear pharmacokinetics.
Animals ; Benzophenanthridines ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Female ; Male ; Rabbits ; Random Allocation
9.Analysis of three wheat cytoplasmic male sterile lines mitochondrial DNA by AFLP.
Qidi ZHU ; Xinbo ZHANG ; M EJAZ ; Gaisheng ZHANG ; Huixue CHE ; Shuping WANG ; Qilu SONG ; Shuling YANG ; Longyu ZHANG
Chinese Journal of Biotechnology 2013;29(5):646-656
Cytoplasmic male sterility is an important way to utilize wheat heterosis. The purpose of thisstudy was to identify cytoplasmic type of three wheat male sterile lines. Amplified fragment length polymorphism (AFLP) marker technique was used to analyze the wheat mitochondrial DNA. We isolated mitochondria by differential centrifugation and density gradient ultracentrifugation. The results show that the extracted mitochondrial DNA was pure. It was suitable for PCR and genetic analysis. We got 4 pairs of specific primers from 64 primers combinations. Primer E1/M7 amplified 3 specific fragments in ms(Kots)-90-110. Primer E4/M2 generated 2 specific fragments in ms(Ven)-90-110. Primer E7/M6 amplified 2 specific fragments in ms(S)-90-110. Primer E6/M4 produced 2 specific fragments in ms(Kots)-90-110. Four specific primers could be used to identify three cytoplasmic types of Aegilops kotschyi, Ae. ventricosa and Triticum spelta. It provided the molecular basis to further study the mechanism of wheat cytoplasmic male sterility.
Amplified Fragment Length Polymorphism Analysis
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methods
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Cytoplasm
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metabolism
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DNA, Mitochondrial
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genetics
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DNA, Plant
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genetics
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Gene Expression Profiling
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Genotype
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Plant Infertility
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genetics
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Triticum
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genetics
10.Study on chromatography-efficacy relation of Zanthoxylum nitidum on gastric cancer cells.
Qingrong SHEN ; Huixue HUANG ; Honghong WANG ; Lijuan CAO ; Kan FENG ; Yuehua YE ; Huagang LIU
China Journal of Chinese Materia Medica 2011;36(19):2693-2696
OBJECTIVETo develop a method for elucidating " chromatography-efficacy" relation of the extract of Zanthoxylum nitidum on the gastric cancer cells.
METHODAfter obtaining the tumor inhibition rate and fingerprint peak data through MTT and HPLC, "chromatography-efficacy" relation was established by an appropriate statistical method.
RESULTThe gastric cancer "chromatography-efficacy" relation of Z. nitidum was established by step-back technique.
CONCLUSIONThe "chromatography-efficacy" relation has statistically significant and practical significance, so it has reference value in some way.
Antineoplastic Agents, Phytogenic ; analysis ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Humans ; Stomach Neoplasms ; drug therapy ; physiopathology ; Zanthoxylum ; chemistry