1.The overexpression of ULBP2 in the brain of drug-refractory epilepsy
Lifeng GUAN ; Xuefeng WANG ; Tao LIN ; Yan ZENG ; Jinmei LI ; Fei XIAO ; Jijun SUN ; Xi ZHU ; Hui YANG ; Guoming LUAN ; Yuping WANG ; Yong LU ; Hongwei LI
Chinese Journal of Immunology 2000;0(11):-
Objective:To study the expression of ULBP2 protein in the brain tissues of patients with drug-refractory epilepsy and its clinical significance.Methods:Gene-chip,immunofluorescence and Western blot were used to test expression of ULBP2 in the surgically removed brain tissue of patients with drug-refractory epilepsy from the brain bank of our department(n=42),and the results were compared with that of normal controls (n=12).Results:The relative increasing expression of ULBP2-gene in the brain of patients with drug-refractory epilepsy,and ULBP2 protein expression was significantly increased in temporal lobe cortex of patients with drug-refractory epilepsy as compared with the same regions of the controls specimens.Conclusion:The results indicate that the overexpression of ULBP2 may be involved in the pathophysiology of drug-refractory epilepsy.
3.Molecular cloning and expression of anti-tumor adhesion peptide (beta3).
Song-Mei WANG ; Jun ZHU ; Yan LI ; Luan-Feng PAN ; Xi-Liang ZHA ; Yin-Kun LIU
Chinese Journal of Biotechnology 2005;21(4):558-562
To block tumor cell adhesion, inhibit tumor metastasis and recurrence, the anti-adhesion peptide-trimeric beta peptide (DLYYLMDLSYSMKGGDLYYLMDLSYSMKGGDLYYLMDLSYSMK, beta3) was designed. The DNA fragment of beta3 was cloned into expression vector pET-His and the fusion protein His-beta3 was expressed in E. coli. BL21(DE3)plysS. After 1.5 hours' induction with IPTG, His-beta3 peptide was expressed significantly amounting to 10% of the insoluble proteins and 4% of the total proteins. 20mg of beta3 peptide was obtained from one litter culture medium after purification by using metal-chelating sepharose 6B FF. The purity of beta3 is 92.2% according to Gel-Pro analysis. The anti-adhesion effects of beta3 peptide, beta1 peptide (DLYYLMDLSYSMK) and GRGDS on the hepatocellular carcinoma cell line SMMC-7721 and the high metastasis hepatocellular carcinoma cell line HCCLM6 were studied. The result showed the beta3 blocked the adhesion of HCCLM6 cells and SMMC-7721 cells to fibronectin (FN) specifically. The inhibition effect was dose-dependent and time-dependent and the inhibition rate of beta3 was higher than three times concentration of beta1 and GRGDS. This suggested that pET-His-beta3/BL21(DE3)plysS was a suitable expression system for beta3, and the expressed beta3 specially inhibited the adhesion of cancer cells.
Amino Acid Sequence
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Antineoplastic Agents
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pharmacology
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Cell Adhesion
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drug effects
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Cloning, Molecular
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Escherichia coli
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genetics
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metabolism
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Molecular Sequence Data
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Peptide Fragments
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pharmacology
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Peptides
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genetics
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metabolism
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pharmacology
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Recombinant Proteins
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biosynthesis
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genetics
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Tumor Cells, Cultured
4.Overexpression of extracelluar signal regulated protein kinase in the temporal lobe of human drug-resistant epilepsy
Zhiqin XI ; Xuefeng WANG ; Yuan WU ; Jijun SUN ; Xi ZHU ; Fei XIAO ; Jinmei LI ; Lifeng GUAN ; Yun GONG ; Fengying LIU ; Yong YAN ; Zhenli GUO ; Guoming LUAN ; Jianguo ZHANG ; Yuping WANG
Chinese Journal of Neurology 2005;0(10):-
Objective To investigate expression of extracelluar signal regulated protein kinase(ERK)and phosphorylation ERK(p-ERK)in the temporal lobe from patients with DR-TLE so as to explore the possible roles of ERK in the pathogenesis of DR-TLE.Methods Expression of ERK was detected with Western blot and immunohistochemistry in 32 patients with DR-TLE(24 temporal lobe,8 hippocampi),as compared with 12 controls(9 temporal lobe,3 hippocampi).Results ERK and p-ERK expression in DR-TLE was significantly higher(0.2266?0.0613,0.2097?0.0183 and 0.1924?0.0054,respectively)than those of controls(0.1840?0.0023,0.1974?0.0056 and 0.1825?0.0063,respectively,all P
5.Clinical characteristics and treatment of Rasmussen syndrome in 16 children.
Yue-hua ZHANG ; Li-hua PU ; Xiao-yan LIU ; Hui XIONG ; Yun-lin LI ; Xing-zhou LIU ; Guo-ming LUAN ; Jiong QIN ; Xi-ru WU
Chinese Journal of Pediatrics 2007;45(9):697-702
OBJECTIVERasmussen syndrome (RS) is a chronic inflammatory disease of unknown origin, usually affecting one brain hemisphere. The present study aimed to analyze the electroclinical characteristics and treatment of RS.
METHODSThe medical records of 16 children with RS were retrospectively reviewed.
RESULTSOf the 16 children, 8 were males and 8 were females. The age of onset was from 1 year and 11 months to 11 years and 6 months. The first symptom was seizure in all patients. The main seizure type was partial motor seizures. In all the patients, seizures gradually became frequent and in the form of epilepsia partialis continua (EPC). Thirteen cases developed hemiparesis. Fixed hemiparesis occurred from 2 months to 3 years after the onset of seizures. The cognitive deterioration was present in 14. The EEG background activity was abnormal in all the cases, asymmetric slow wave disturbances were bilateral but with unilateral predominance in 11, unilateral delta or theta wave in 8. The presence of interictal epileptiform discharges were found in all cases, unilateral in 11 and bilateral in 5. Seizures were recorded in all patients, no electroclinical correlation was found in 5. Serial magnetic resonance imaging (MRI) showed progressive unihemispheric or focal cortical atrophy in all cases. Six cases transiently showed focal cortical swelling or T2/FLAIR hyperintense signal on early scans. Antiepileptic drugs were not effective in any of the patients. Three of 10 patients receiving immunoglobulin, and 4 of 8 receiving corticosteroids, had some reduction of seizure frequency for a short period. Six patients accepted functional hemispherectomy, in 4 of them seizure no longer occurred and cognitive function was improved. The results of multiple subpial transection in 2 cases and focal resection in one patient were disappointing.
CONCLUSIONThe clinical features of RS were refractory partial epilepsy, progressive hemiplegia and cognitive deterioration. The EEG background was asymmetric with slow wave activity, interictal epileptiform discharges were unilateral or bilateral, no electroclinical correlation occurred. Serial MRI showed progressive unihemispheric focal cortical atrophy. Antiepileptic drugs were not effective for RS. In some patients, immunoglobulin or corticosteroids could reduce seizure frequency in the short term. Functional hemispherectomy could lead to seizure control and prevent further development of neurological impairment and cognitive deterioration.
Anticonvulsants ; therapeutic use ; Child ; Child, Preschool ; Cognition ; drug effects ; Electroencephalography ; Encephalitis ; drug therapy ; physiopathology ; prevention & control ; Epilepsia Partialis Continua ; drug therapy ; prevention & control ; Epilepsy ; etiology ; prevention & control ; Female ; Hemispherectomy ; methods ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Treatment Outcome
6.Study on UGT1 A1?6 gene polymorphisms in cancer patients of Han population in Anhui area
Jia-Jie LUAN ; Jun LIU ; Lin WANG ; Yan-Hong ZHU ; De-Xi ZHOU ; Jian ZUO ; Zhen-Yu XU
Chinese Pharmacological Bulletin 2018;34(6):857-862
Aim To investigate the gene frequency of UGT1 A1?6 in cancer patients in Anhui Han popula-tion. Methods The 222 cases of blood samples of Han cancer patients were collected from different re-gions of Anhui province, and the UGT1A1?6 geno-types were detected by in situ hybridization fluorescencestaining. Results Patients with a UGT1A1?6 wild type ( GG ) accounted for ( 159 cases, 71. 62%) , which were higher than those of heterozygous mutations ( GA, 52 cases, 23. 42%) and of homozygous muta-tions ( AA, 11 cases, 4. 96%) of the total cases. The mutation rate of UGT1 A1?6 was 16. 67%, and partic-ularly in patients with esophageal cancer it was 43. 75%. The rates of mutation in the patients in Ma ' anshan and Chuzhou were 40. 01% and 34. 62%, re-spectively, both significantly higher than those of othertumors and regions. Conclusions Cancer patients in Anhui Han population have a high mutant frequency of UGT1 A1?6 . The UGT1 A1?6 genotyping can indi-rectly predict the risk of irinotecan's adverse reaction, which obviously enhances the potentially individualized treatment of irinotecan.
7.Cloning and expression of Streptococcus salivarius urease gene in Escherichia coli
Yan WANG ; Xi-Ping FENG ; You-Hua XIE ; Dan-Ying TAO ; Xiao-Ling LUAN
Chinese Journal of Stomatology 2010;45(8):498-501
Objective To clone Streptococcus salivarius(Ss) 57. I urease gene, which can express ureolytic activity in Escherichia coli (Ec) without adding extra nickel ions. Methods Urease gene was cloned by polymerase chain reaction in three separate parts. The three separate plasmids were digested by specific restriction enzymes and ligated together. The expression of the complete urease gene in Ec was detected by phenol red assay and pH analysis. Results Urease gene of Ss 57. I was eventually cloned and proved correct. Urease activity of the obtained clone was positive in Ec. Without adding extra NiCl2, the recombinant Ec could hydrolyze urea to produce ammonia, resulting in the increase of pH value.Conclusions The clone of Ss urease gene obtained in this study could express ureolytic activity in Ec without adding extra nickel ions. The current clone can be used to construct ureolytic effector strain used in replacement therapy in caries prevention.
8.Quantitative Detection of Procalcitonin in Blood by Nanozyme-based Lateral Flow Immunoassay
Yue ZHENG ; Tong LIN ; Yong-Hua XIONG ; Meng-Shuo XU ; Xi-Luan YAN ; Xu-Jing GUO ; Lei YANG ; Liang GUO
Chinese Journal of Analytical Chemistry 2024;52(8):1082-1093
A rapid quantitative immunochromatographic assay for procalcitonin(PCT)using metal-organic frameworks modified with gold and platinum nanoparticles(MAPs)as labels was established in this work.The detection probe was prepared by conjugating MAPs with anti-PCT monoclonal antibody via an electrostatic adsorption method.Anti-PCT polyclonal antibody and sheep anti-mouse IgG were sprayed onto the nitrocellulose(NC)membrane as the test line and quality control line,respectively,to construct immunochromatographic strip for PCT quantitative detection via signal-amplification-based sandwich immunoassay.The results showed that the MAP-based immunochromatographic test had high sensitivity,high specificity,and good stability.The dynamic range for detection of PCT was 0.61 pg/mL-320 ng/mL,the detection limit was 0.25 pg/mL,and the intra-day and inter-day precision(Relative standard deviation)were less than 15%.The results of real sample analysis showed that a quite low volume of sample was required for detection of PCT in whole blood,which was of great significance for the early diagnosis,monitoring and treatment,and prognosis of inflammation.
9.Effects of Senegenin against hypoxia/reoxygenation-induced injury in PC12 cells.
Xiao-Qing ZHU ; Xue-Min LI ; Yan-Dong ZHAO ; Xi-Luan JI ; Yan-Ping WANG ; Yong-Mei FU ; Hua-Dong WANG ; Da-Xiang LU ; Ren-Bin QI
Chinese journal of integrative medicine 2016;22(5):353-361
OBJECTIVETo investigate the effect and the potential mechanism of Senegenin (Sen) against injury induced by hypoxia/reoxygenation (H/R) in highly differentiated PC12 cells.
METHODSThe cultured PC12 cells were treated with H/R in the presence or absence of Sen (60 μmol/L). Four groups were included in the experiment: control group, H/R group, H/R+Sen group and Sen group. Cell viability of each group and the level of lactate dehydrogenase (LDH) in culture medium were detected for the pharmacological effect of Sen. Hoechst 33258 staining and annexin V/propidium iodide double staining were used to analyze the apoptosis rate. Moreover, mitochondrial membrane potential (△Ψm), reactive oxygen species (ROS) and intracellular free calcium ([Ca(2+)]i) were measured by fluorescent staining and flow cytometry. Cleaved caspase-3 and activity of NADPH oxidase (NOX) were determined by colorimetric protease assay and enzyme linked immunosorbent assay, respectively.
RESULTSSen significantly elevated cell viability (P<0.05), decreased the leakage of LDH (P<0.05) and apoptosis rate (P<0.05) in H/R-injured PC12 cells. Sen maintained the value of △Ψm (P<0.05) and suppressed the activity of caspase-3 (P<0.05). Moreover, Sen reduced ROS accumulation P<0.05) and [Ca(2+)]i increment (P<0.05) by inhibiting the activity of NOX (P<0.05).
CONCLUSIONSen may exert cytoprotection against H/R injury by decreasing the levels of intracellular ROS and [Ca(2+)]i, thereby suppressing the mitochondrial pathway of cellular apoptosis.
Animals ; Apoptosis ; drug effects ; Calcium ; metabolism ; Caspase 3 ; metabolism ; Cell Hypoxia ; drug effects ; Cell Nucleus ; drug effects ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Fluorescence ; Intracellular Space ; metabolism ; Membrane Potential, Mitochondrial ; drug effects ; NADPH Oxidases ; metabolism ; Neuroprotective Agents ; pharmacology ; Oxygen ; pharmacology ; PC12 Cells ; Rats ; Reactive Oxygen Species ; metabolism ; Staining and Labeling
10.History of atopy reduces predictive value of fractional exhaled nitric oxide for eosinophilic airway inflammation in chronic cough.
Yan Mei YE ; Da Peng HU ; Shu Yu HUANG ; Wen Qu ZHAO ; Jian Peng LIANG ; Jun Wen HUANG ; Hui Shan HAN ; Shu Luan YANG ; Shao Xi CAI ; Hai Jin ZHAO
Journal of Southern Medical University 2022;42(10):1470-1475
OBJECTIVE:
To investigate the impact of a history of atopy on the value of fractional exhaled nitric oxide (FENO) for predicting sputum eosinophils in patients with chronic cough.
METHODS:
A total of 868 patients with persistent cough lasting more than 3 weeks without pulmonary infection were enrolled, including 119 patients with subacute cough (defined as cough lasting 3-8 weeks) and 749 with chronic cough (longer than 8 weeks). The predictive value of FENO level for sputum eosinophilia was analyzed using receiver-operating characteristic (ROC) curve analysis, and the area under the curve (AUC) was calculated. The atopy status of the patients was determined by screening for history of allergy, hay fever, or animal or food allergies.
RESULTS:
Of the 868 patients enrolled, 173 patients (19.9%) had eosinophilic airway inflammation (EAI). In the overall patients, the median (Q1, Q3) FENO level was 18 (12, 35) ppb, ranging from 5 to 300 ppb. The patients with chronic cough and a positive history of atopy had a higher median FENO level than those without atopy (24 [13, 50] vs 18 [11, 34]; Z=2.25, P= 0.029), and FENO level was significantly correlated with EAI (r=0.281, P < 0.001). The AUCs of FENO for diagnosis of airway eosinophilia in patients with atopy and those without atopy were 0.677 (95% CI: 0.548-0.806) and 0.708 (95% CI: 0.660-0.756), respectively. The optimal cut-off value of FENO for diagnosing EAI was higher in patients with atopy than in those without atopy (72 vs 28.5 ppb).
CONCLUSION
A history of atopy reduces the predictive value of FENO level for EAI in patients with chronic cough, suggesting the importance of examining the atopic status when interpreting test results of FENO.
Humans
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Cough/diagnosis*
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Exhalation
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Fractional Exhaled Nitric Oxide Testing
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Nitric Oxide/analysis*
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Eosinophilia
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Chronic Disease
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Inflammation