1.Effect of different doses of sinomenine on CD80 and CD86 expressions and extracellular interleukin-12 secretion in dendritic cells
Chinese Journal of Tissue Engineering Research 2007;11(28):5654-5656
BACKGROUND: Sinomenine, an alkaloid monomer extracted from Chinese medicinal herb sinomenium acutum,possesses potent anti-inflammatory, analgesic and immunoinhibitory pharmacological activities. Sinomenine has certain therapeutic effect on rheumatoid arthritis and has been widely applied in the clinic. Dendritic cell (DC) is the known antigen presenting cell with the strongest functions in the body.OBJECTIVE: To observe the effect of different doses of sinomenine on CD80 and CD 86 expression and extracellular interleukin-12 secretion in DCs.DESIGN: A controlled repeated measuring study based on the cells.SETTTNG: Department of Pharmacology, Guangdong Medical College.MATERIALS: The experiment was carried out in the Institute of Immunology, Third Military Medical University of Chinese PLA between September 2004 and May 2005. The peripheral blood of healthy donors was supplied by the blood bank of the Southwest Hospital. RPMI1640 medium was purchased from Hyclone Company. Fluorescein isothiocyanate(FITC)-conjugated mouse monoclonal antibodies against CD80 and CD86 and FITC-conjugated mouse IgG1 were purchased from Bioscience Company. Recombinant human interleukin-4 (rhlL-4), recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and tumor necrosis factor-α (TNF-α) were purchased from PharMingen Company. Sinomenine was supplied by Zhengqing Pharmacy Company in Hunan. ELISA kit specific for IL-12 was purchased from Diaclone Company.METHODS: Peripheral blood was collected from healthy volunteers. 1 ×106 U/L recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and 5×105 U/Lrecombinant human interleukin-4 (rhlL-4)were added to the peripheral blood. After 7 days, DCs were harvested. ①The DCs were counted and cultured at 2×108 L 1 in fresh medium supplemented with TNF-α and different doses of sinomenine(3, 10, 30 mg/L). After 48 hours, 2×105 cells were collected and washed twice with phosphate buffer solution (PBS). Then the expressions of CD80 and CD86 were analyzed using 20 μL FITC-conjugated mouse monoclone antibodies against CD80 or CD86.FITC-conjugated mouse IgG1 was used as control at 4 ℃ away from light for 30 minutes. After staining, cells were washed twice with PBS and fixed in 10 g/L paraformaldehyde. The samples were analyzed on a flow cytometry. ②The DCs were counted and cultured at 2 ×108 L-1 in fresh medium supplemented with TNF-α and different doses of sinomenine (3, 10, 30 mg/L). After 48 hours, the supernatants were collected and the concentration of interleukin-12 (IL-12) was determined in a sandwich ELISA using kit specific for IL-12 according to the manufacture's instruction.MAIN OUTCOME MEASURES: ①Effect of different doses of sinomenine on the expressions of CD80 and CD86 on DCs. ②Effect of different doses of sinomenine on the level of IL-12 secreted by DCs.RESULTS : ① Expressions of CD80 and CD86 on DCs: There were no significant differences in the percentage of cells positive and fluorescence intensity for CD80 and CD86 on DC between different doses of sinomenine groups and control groups (P > 0.05). ② IL-12 level secreted by DCs: From ELISA measurement, we found that IL-12 level of the sinomenine 3 mg/L, 10 mg/L and 30 mg/L groups was (863.1±33.7), (668.8±31.9), (512.6±29.6) ng/L, respectively,which was higher than that of control group [ (922.2±36.6),P < 0.05-0.01]. The level of IL-12 in the supernatants of DCs co-cultured with sinomenine was significantly lower compared with control, which was dose-dependent.CONCLUSTON: Sinomenine can inhibit the level of IL-12 secreted by DCs in dose-dependent manner, but do not influence the expressions of CD80 and CD86 in DCs; The therapeutic effect of sinomenine on rheumatoid arthritis may be related to its inhibition to secretion of IL-12 in DCs.
2.Progress of cathepsin D in cancer
International Journal of Surgery 2012;39(3):192-196
Tumor cell invasion and metastasis are associated with the proteolytic activity of various types of proteinases.Among them,cathepsin D,which is a lysosomal proteinase,has received more attention recently.Various studies have shown that the lysosomal aspartic protease cathepsin D is over-expressed and hyper-secreted by numerous cancer cell lines.Indeed it plays an essential role in the multiple steps of tumor progression,in stimulating cancer cell proliferation,tumor invasion and metastasis,fibroblast outgrowth and angiogenesis,as well as in inhibiting tumor apoptosis.In addition,CD is also a key mediator of induced- apoptosis.The aim of this article is to review the current knowledge on Cathepsin D action in cancer progression and metastasis,as well as its dual function in apoptosis.
3.Practice and exploration of emergency clinical training for medical students
Chinese Journal of Medical Education Research 2012;11(4):394-396
Objective To compare the results of emergency clinical training for medical students and to put forward some suggestions.Methods Toatlly 207 medical students of Grade 4 studied in the first people' s hospital of Shanghai Jiaotong university from 2009 to 2011 were divided into the traditional group (n =90 ) and the experimental group (n =117 ).The students in the experimental group participated in emergency clinical training in summer vacation while those in the traditional group did not.Questionnaires about their self-learning ability,clinical communicating ability and critical thinking ability after training the were conducted and the data were analyzed by descriptive statistical analysis.Results One hundred and five effective questionnaires were recovered from the experimental group,with the recovery rate 89.7% and 62 effective questionnaires were recovered from the traditional group,with the recovery rate 68.9%.The scores about clinical communicating ability and critical thinking ability of the experimental group were significantly highcr than those of the traditional group [ (79.45 ± 9.354)vs.(75.87 ± 9.926),(263.38 ± 30.925 ) vs.(251.36 ± 23.679),P < 0.05 ].However,there was no significant difference in scores about self-learning ability between the two groups [ ( 153.97 ± 23.725 ) vs.( 149.83 ± 13.891 ),P > 0.05].Conclusion Emergency clinical training is helpful to improve medical students' clinical communicating ability and critical thinking ability.It is worth popularizing.
4.Investigation on the rusuits of implementing the tutorial system for medical interns
Chinese Journal of Medical Education Research 2011;10(6):742-746
ObjectiveTo analyze the results of implementing the tutorial system for medical interns, and to put forward some suggestions. MethodsThe medical interns and the doetors selected as tutors were asked to do questionnaires to study the tutorial system's feasibility and effectiveness. Simultaneously, we adopted the Objective Structured Clinical Examination ( OSCE ) to assess its process and resuits. Results74.3% of the students and 75.5% of the doctors think that they need the tutorial system,73.2%of the students think that this system can train better clinical skills,67.9% of the doetors think that they can review the knowledge learned through teaching. Implemented after one year, students' OSCE scores are improved. ConclusionTutorial system for medical interns has achieved preliminary success, and the preparatory work and related-system building need to be further improved.
5.The relationship among personality,job burnout and knowledge sharing behavior of the enterprise employees
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(8):722-724
Objective To explore the relationship among personality,job burnout and knowledge sharing behavior of the enterprise employees.Methods The 44 items big five inventory(BFI) ,knowledge sharing inventory and job burnout inventory were administrated to 635 employees online.Results ( 1 ) Agreeableness had a significant main effect on knowledge sharing ( β =0.135, P<0.01); (2)Cynicism( β = -0.140, P<0.01) and reduced personal accomplishment( β = -0.125, P<0.01 ) had a significant effect on knowledge sharing behavior and its two dimensions; (3) Job burnout partially mediated the relationship between agreeableness and knowledge sharing behavior(P<0.01).Conclusion Enterprise employees' personality had different effects on knowledge sharing behavior,and job burnout was an important mediating role in the relationship above.
6.Experimental study on edaravone in prevention of restenosis in rat common carotid balloon angioplasty
Chinese Journal of Neurology 2008;41(9):633-637
Objective To investigate edaravane in prevention of restenosis in rat common catery arteries balloon angioplasty and the possible mechanism of this process. Methods Forty male SD rats were randomly divided into 2 groups: treatment group (whose intima was injured by balloon and was given edaravone 3 mg/kg by peritoneal injection, bid) and control group (whose intima was injured by ballon and was given the same volume NS as edaravone). 1, 3, 7, 14 and 28 days after angioplasty, rats were killed and the local arteries were sectioned for analysis of pathological morphology and the expression of matrix metalloproteinase-2 (MMP-2) analyzed by immunohistochemistry. Results Small amounts of smooth muscle cell appeared at the intima of blood vessel 3 days after the artery injured by balloon. Consecutive intima was formed 7 days after artery injury. The intima was even or uneven thickened and the lumen of artery was significant stenosed 14 and 28 days afterwards. In the control group, the intimal area reached (0.240±0.043) mm2, the intima hyperplasy index were 0.52±0.06, stenosis ratio were 30%±9% 14 days afterwards. The intimal area were (0.420±0.063 )mm2, the intima hyperplasy index were 0.67± 0.07, stenosis ratio were 54%±9% on day 28. In the treatment group, the intimal area were (0.063± 0.025)mm2, the intima hyperplasy index were 0.24±0.07, stenosis ratio were 8%±3% after 14 days. The intimal area were (0.116±0.023) mm2, the intima hyperplasy index were 0.38±0.05, stenosis ratio were 16%±4% after 28 days. Compared with control group at the same time, the intimal area (F values were 50.488 and 81.119 respectively, P < 0.05 ), the intima hyperplasy index ( F values were 41.743 and 48.122 respectively, P<0.05) and stenosis ratio (F values were 24.221 and 81.119 respectively, P< 0.05) were decreased in treatment group. The expressions of MMP-2 raised after balloon injury. In control group, the expressions of MMP-2 were 27.16%±7.15% after 3 days, 22.59%±6.68% after 7 days, 18.85%±4.91% after 14 days. In the treatment group, it decreased to 15.57%±3.62% after 3 days, 12.91%±1.88% after 7 days. Compared with control group, the expressions of MMP-2 in treatment group were lower after 3 days and 7 days ( F = 8.359, P = 0.028 and F = 7.781, P = 0.032 ). Conclusions Edaravone can prevent restenosis in rat common catery after balloon angioplasty. The possible mechanism is that edaravone can inhibit the expressions of MMP-2 in this process.
7.Clinical and electroencephalographic characteristics of frontal lobe epilepsy
Clinical Medicine of China 2008;24(7):676-677
Objective To explore the clinical and electroencephalographic characteristics of frontal lobe ep- ilepsy(FLE).Methods The clinical and electroencephalographic data of 80 patients with FLE were analyzed.Re- suits The seizure types of frontal lobe epilepsy were simple focal seizure-complex focal seizure and secondary gen- eral tonic.clonic seizure.Seizures were often exhibited in relatively short duration,hyper motor,tonic or postural.Fre- quent nocturnal attacks and slight postictal mental confusion was increased.Interictal EEG manifested sharps or spikes or slow wave in the frontal area.Ictal EEG showed paroxysm rhythms.Conclusion FLE is a distinct epilepsy syndrome.
8.Construction and identification of luciferase reporter gene containing mouse T-bet promoter.
Chinese Journal of Biotechnology 2014;30(11):1733-1741
The aim of this study is to clone the mouse T-bet promoter and enhancer, construct and identify the firefly luciferase reporter gene plasmid pGL4.10-TBX21pr-CNS for T-bet transcription regulation study and its function in signaling of multiple sclerosis. The promoter and CNS of T-bet were predicted by bioinformatics assay. The predicted fragment of mouse T-bet promoter plus CNS was amplified by PCR and cloned into pGL4.10. The recombinant plasmid pGL4.10-TBX21pr-CNS was transferred into Escherichia coli DH5α. The positive clone was identified by double digestion with Kpn I and Sfi I and DNA sequencing. Finally, pGL4.10-TBX21pr-CNS was cotransfected with pRL-TK into 293T cells and Jurkat cells, pRL-TK and pGL4.10 as a control. The luciferase activity in 293T cells (P = 0.012 2) and Jurkat cells (P = 0.002 2) was higher than that of the control group. A fragment of 1 028 bp mouse T-bet promoter plus 1 308 bp CNS was successfully cloned and the firefly luciferase reporter gene plasmid pGL4.10-TBX21pr-CNS was constructed. In 293T cells and Jurkat cells, pGL4.10-TBX21pr-CNS has the promoter functions. This work offers a basic material for the research of T-bet transcription.
Animals
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Gene Expression Regulation
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Genes, Reporter
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Genetic Vectors
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Luciferases
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Mice
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Multiple Sclerosis
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Plasmids
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Promoter Regions, Genetic
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T-Box Domain Proteins
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genetics
9.Effect of intensive blood pressure reduction on prognosis and early hematoma growth in acute intracerebral haemorrhage
Chinese Journal of Emergency Medicine 2010;19(12):1253-1257
Objective To study the safety and efficacy of intensive reduction of blood pressure for the treatment of acute cerebral haemorrhage. Method A randomized control trial in 41 consecutive patients with intracerebral haemorrhage admitted from October 2006 to January 2007 were randomly assigned to intensive blood pressure reduction group ( n = 24) or guidelines blood pressure reduction group ( n = 17) (tho guidelines set by American Association of cardiologists). In the intensive reduction group, the systolic pressure was reduced immediately to lower than 140 mmHg, while the blood pressure was reduced to that just below 180 mmHg in guideline reduction group. The size of the haematoma was measured 24 h after treatment by CT scans and the patients were followed up for 90 days. Death and/or disability in 90 days, and the short-term and long-term neurological function and the size of haematoma in 24 hours of two groups were compared. The outcomes were statistically analyzed with SPSS version 10.0 software. Measurement data were analyzed with t -test while numeration data were analyzed with chisquare test. Results There were no significant differences either in death and/or disability or in short-term and long-term neurological function in 90 days after treatment ( P > 0.05). The mean values of proportional enlargement of haematoma were 16.8% in the intensive group and 36. 1% in the guidelines group 24 hours after treatment ( P = 0.012). The mean values of absolute enlargement of haematoma of two groups were 2.7 mL and 5. 1 mL,respectively (P = 0.058). There was significant difference in rate of enlargement of haematoma in the early stage of acute cerebral haemorrhage (4.2% vs. 47. 1%, P = 0.012). Conclusions Although intensive reduction of blood pressure in patients with acute cerebral haemorrhage did not alter the clinical prognosis of patients, it could apparently attenuate the enlargement of haematoma in the early stage of acute cerebral haemorrhage.
10.Comparative efficacy of intra-arterial thrombolysis with urokinase and intravenous thrombolysis with recombinant tissue plasminogen activator for acute ischemic stroke
International Journal of Cerebrovascular Diseases 2010;18(10):726-729
Objective To evaluate the efficacy and safety of intra-arterial thrombolysis with urokinase and intravenous thrombolysis with recombinant tissue plasminogen activator (rtPA) for acute ischemic stroke. Methods A total of 43 patients with acute ischemic stroke within 6 hours of onset were included, 31 of them underwent superselective intra-arterial thrombolysis in the intra-arterial thrombolysis group and 12 of them underwent intravenous thrombolysis with rtPA in the intravenous thrombolysis group. Vascular recanalization was observed in the intra-arterial thrombolysis group, and the modified Rankin scale (mRS) scores at day 90 were used to evaluate the outcomes in both groups. Results Eighteen patients (58.1%) had complete recanalization and 7 (22.6%) had partial recanalization in the intra-arterial thrombolysis group. The recanalization rate was 80.6%, 3 complicated symptomatic intracranial hemorrhage, and 1 died. There vere no significant differences between the good outcome rate (74.2% vs. 66.7%, x2 =0.24, P=0.622) and the incidence of symptomatic intracranial hemorrhage at 90 d (9. 68% vs. 8. 33%, x2 =0. 19, P =0. 892). Conclusions Urokinase intra arterial thrombolysis within the time window can significantly improve the recanalization rate of the occluded vessels and improve the clinical symptoms of the patents in acute phase and long term outcomes. Their short-term efficacy and long-term outcomes are almost the same with intrave nous thrombolysis with rtPA.