1.Development of SMART Technology and It’s Application
China Biotechnology 2006;0(06):-
Switching Mechanism At 5′ end of the RNA Transcript(SMART)is a technology used in biology researching,So far,there is no review only about SMART technology.So,the aim is to investigate the research developments of principles,methods and applications of the SMART technology.Based on some researches and combined with the review of the related literature at home and abroad,it analyzed and evaluated the latest development of the research on SMART.As the applications of the SMART technology expand in many fields day after day,it has been proved that the SMART technology is a very useful and efficient skill to construct full length cDNA library.As more and more researchers know this technology,the advantages of the SMART technology become obvious,and meanwhile the disadvantages of the SMART technology also show up.That is to say,the technology needed to be improved.
2.Discussion of MRI segmentation by using FCM
Wei DOU ; Xin HUANG ; Weisong YANG
Chinese Journal of Medical Physics 2000;17(4):201-202
In this paper, a segmentation method, supervised FCM, is used to segment multi-spectrum MR imaging. The qualitative evaluation of human brain can be provided by the results for diagnostics. It can improve the results using FCM.
3.The apoptosis-inducing effect of MnSOD_m on K562 cells and its molecular mechanism
Linlan FAN ; Hulai WEI ; Wei DOU ; Weisheng LIU ; Huifang ZHANG
Chinese Pharmacological Bulletin 2003;0(10):-
Aim To explore the apoptotic effect of mimics of manganese superoxide dismutase(MnSODm)on human leukemia cell line K562 in vitro and the possible molecular mechanisms.Methods Human leukemia K562 cells were used as the target cells.The cell proliferating activity was examined by a MTT colorimetric assay,and the apoptosis of K562 cells was assessed with FITC-Annexin V and propidium iodide(PI)double staining and morphological changes.The expressions of bcl-2 and bax mRNA were detected by reverse transcription polymerase chain reaction(RT-PCR),and flow cytometry(FCM)was employed to measure the expressions of Bcl-2 and Bax protein,mitochondrial inner membrane potential(??m),Cytochrome C(Cyt C)release and Caspase-3 activity.Results The proliferation of K562 cells was obviously inhibited by 0.5~10 mg?L-1 MnSODm(P
4.The clinical characteristics of patients with thymoma-associated myasthenia gravis
Yuping CHEN ; Wei WANG ; Zhongkui WANG ; Yunke DOU ; Dongning WEI
Chinese Journal of Internal Medicine 2012;51(8):623-625
Objective To investigation the clinical characteristics in myasthenia gravis (MG)patients with thymomas.Methods A total of 856 MG patients admitted to the department during 2008.7-2010.12 were reviewed retrospectively.The patients with MG were divided into two groups based on thymic pathology,which were 162 cases with thymoma and 694 cases without thymoma.We compared the different clinical features including the gender,age of onset,MG symptoms and the incidence rate of myasthenia crisis.And the relationship between the WHO types,Maosaoka stages of thymoma and the severe of MG was also studied.Results The percentage of thymoma-associated MG patients was 18.9 percent of hospitalized MG patients at the same period.Of the 162 thymoma-associated patients,94 were male and 68 were female,with a ratio of 1.38∶1 and a mean age of (42.9 ± 12.4)years old.Thymoma was more frequent in middle-old aged patients than in children.Compared with non-thymoma MG,more thymomatous patients showed generalised MG,but not only ocular muscles weakness (90.1% vs 62.4%,P < 0.001 ).There were significant differences of the incidence rate of myasthenic crisis in the two groups ( 14.8% vs 2.3% ).(2)WHO type B2 and Maosaoka Ⅰ,Ⅱ thymoma were the commonest types among all potentially MG-associated thymoma.No differences of Osserman MG classification was found in thymomatous patients with different pathologic changes.Conclusions The thymomatous MG patients had its distinctive clinical features:thymomas occured in about 19.7% of MG patients with more men than women,more common in generalized,higher incidence of myasthenia crisis,with B2 type thymic pathology and Maosaoka Ⅰ,Ⅱstages.No correlation was found between pathologic and clinical stagcs.
5.The effect of sodium 4',7-bihydroxylisoflavone-sulfonate on gastric motility and its mechanism in rat.
Wei LI ; Shu-Cheng AN ; Dou-Dou LUO
Chinese Journal of Applied Physiology 2007;23(1):106-110
AIMTo explore the effect of sodium 4',7-bihydroxylisoflavone-sulfonate (SBIS) on gastric motility in rats and to analyse its mechanisms.
METHODSUsing intraperitoneal (ip) injection and intracerebroventriular (icv) microinjection of drugs and recording the frequency and amplitude of contraction of gastric motility.
RESULTS(1) The injection (ip) of different doses of SBIS could decrease the amplitude of gastric motility, but it wasn't a dose-dependent effect. SBIS also had no effect on the frequency of contraction. (2) The ip injection of naloxone reversed the inhibitory effect of SBIS on the amplitude of gastric contraction. (3) The effect of SBIS could be increased by the ip injection of propranolol and be reversed by the ip injection of phentolamine. (4) After the ip injection of atropine, the effect of SBIS on gastric motility had not been changed remarkably. (5) Different doses of SBIS had been microinjected (icy), but only the small dose decreased the amplitude of gastric motility and also the frequency of contraction had not been markedly changed.
CONCLUSIONBoth the i.p. and icv injection of SBIS can inhibit the gastric motility. Its effect can be achieved at least not only by endogenous opioid peptide and its receptors, but also adrenergic neuron and its alpha-receptors. Adrenergic neuron and its beta-receptors are also involved in the modulating effect of SBIS.
Animals ; Gastrointestinal Motility ; drug effects ; Isoflavones ; pharmacology ; Male ; Rats ; Rats, Sprague-Dawley
6.Effect of RhoA gene silencing on proliferation and migration of HepG2 cells
Wei DONG ; Kefeng DOU ; Xuekang YANG ; Fuqin ZHANG ; Desheng WANG
Chinese Journal of Digestive Surgery 2010;9(3):216-219
Objective To construct a RhoA-siRNA expression vector and determine its role on the malig-nant behavior of HepG2 cells.Methods A RhoA-siRNA DNA fragment was synthesized and cloned into the expression vector of pGenesil-1.The constructed Rhon-siRNA DNA plasmid was stably transfected into HerG2 cells by lipofectamine,and then HepG2 cells were divided into the HepG2/RhoA-siRNA group (HepG2 cells were transfected with pGenesil-1-RhoA-siRNA),HepG2/control group(HepG2 cells were transfected with control plasmid) and HepG2 group (without plasmid transfection).The inbibitory effect of RhoA-siRNA on RhoA protein expression was shown by Western blot.The proliferation,migration,growth potentiality and cell cycle of transfected HepG2 cells were evaluated by MTT assay,wounded healing,the plate cloning formation test and flow cytometry,respectively.All data were analyzed by one-way analysis of variance (ANOVA) and chi-square test.Results The expression of RhoA protein in the HepG2/RhoA-siRNA group was,significantly decreased compared with that in the other two groups (F=178.19,P<0.05).Scratched cells were healed within 48 hours in the HepG2/control group and HepG2 group,but not in the HepG2/RhoA-siRNA group.The clone formation rates in the HepG2/RhoA-siRNA group,HepG2 group and HepG2/control group were 39%±3%,67%±5%and 70%±6%,respectively,with a significant difference among the three groups(χ2=33.34,38.69,P<0.05).Flow cytometry showed that the number of cells transfected with RhoA-siRNA was highest in the G0/G1 phase and lowest in the S phase(F=70.46,76.57.P<0.05).Conclusion The RhoA-siRNA expression vector can effectively suppress the proliferation and migration of HepG2 cells,which may provide a novel gene therapy for hepatocellular carcinoma.
7.Effects of yichan tang on rotatory behavior, substance nigra cells and neurotransmitter of rats with Parkinson disease
Yongqi DOU ; Minghui YANG ; Wei WANG ; Yi LIU
Chinese Journal of Tissue Engineering Research 2006;10(31):168-171
BACKGROUND: Parkinson disease (PD) not only influences motor function, also affects sensation and cognition. Treatment with dopamine can only control symptom and has well side effect; however, yichan tang has great effect on treating PD in clinic.OBJECTIVE: To observe the effect of yichan tang on praxiological behavior, catecholamines substance in cerebral tissue and substantia nigra nerve cells of PD rats and analyze its effective mechanism.DESIGN: Randomized control animal study and single-blind evaluation.SETTING: Department of Traditional Chinese Medicine of General Hospital of Chinese PLA.MATERIALS: A total of 120 female SD rats weighing 180-200 g were selected in this study. Seventy modeling PD rats were randomly divided into saline group (n=30) and yichan tang group (n=40) according to digital table. Another 30 non-modeling rats were regarded as normal control group. Yichan tang was made of 15 g shanyurou, 15 g shichangpu, 10 g xianlingpi, 10 g roucongrong, 15 g gouqizi, 15 g danshen and 8 g wugong,etc., and provided by Dispensary of Chinese herb of General Hospital of Chinese PLA. Drug was decocted for 30 minutes to filter the solution and then concentrated into decoction including 1.0 g/mL raw materials.METHODS: The experiment was carried out at the Animal Laboratory of General Hospital of Chinese PLA from September to December 2001. PD rat model were established with 6-hydroxy-dopa to observe behavior of rats.The administration of rats (body mass/kg) in yichan tang group was 10 times as the administration of the adults (60 kg) (5 g/kg). Rats were perfused once a day and weighted once a week. The dosage of administration was regulated as the body mass changing, and the administration lasted for 8 weeks. Rats in saline group and normal control group were perfused with the same volume of saline as the criteria of body mass. Rotatory behaviors of rats in normal group, saline group (model group before administration)and yichan tang group were observed before administration and at 5, 6, 7 and 8 weeks after administration. Changes of content of catecholamines in extracellular fluid of cerebral tissue were measured with micro-dialysis technique and high performance liquid chromatography (HPLC), meanwhile, changes of substantia nigra nerve cells on both sides of corpus striatum were observed under 20-time and 40-time optic microscopes.MAIN OUTCOME MEASURES: Effect of yichan tang on medium of cranial nerve of PD rats; changes of substantia nigra nerve cells on both sides of corpus striatum with optic microscope and electron microscope; evaluation of yichan tang.RESULTS: Among 100 rats, 6 rats in yichan tang group died because of failure perfusion, 5 in saline group and 2 in normal control group. Therefore, 87 rats were involved in the final analysis. ① At 5, 6, 7 and 8 weeks after treatment, rotatory times within 40 minutes were all shorter in yichan tang group than those in saline group [(112.34±33.36), (91.16±73.06),(72.05±20.77), (61.63±17.93), (340.90±52.97) times, P < 0.01]. ② Levels of 3,4-dihydroxybenzene acid, homovanilic acid, dopamine and 5-serotonin on the injured side were lower than those on the uninjured side (P < 0.05-0.01). After treatment, levels of mediums mentioned above were higher in yichan tang group than those in saline group (P < 0.05-0.01); however,there were no significant differences of mediums on the uninjured side in the three groups (P > 0.05). ③ Pathological observation: Amount of substantia nigra nerve cells on the injured side was more in yichan tang group than that in saline group; moreover, volume of neurons was full, structure was clear, and values of Golgi complex and mitochondria were close to the normals.CONCLUSION: Yichan tang can relieve injured degrees of substantia nigra nerve cells, facilitate the repair, increase the content of endogenetic catecholamines in cerebral tissue, and improve rotatory behavior of PD rats.
8."""Exploring the Unknown, the Pursuit of New Knowledge"", Improving the Level of Chinese Medicine Academic and the Modern Basic Theory of Traditional Chinese Medicine Research---Series of Studies on the Modern Basic Theory of Traditional Chinese Medicine (Part 2-II)"
Mingqi QIAO ; Xianghong ZHAN ; Xuejun DOU ; Sheng WEI ; Xiaoru WANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(7):1442-1450
To guarantee the research and establishment of the modern basic theory of traditional Chinese medicine, We firstly proposed the concept of the academic standards of the modern basic theory of traditional Chinese medicine and its logical structure and standards points in this paper. According to the research progress at home and abroad, we explored the basic concept of academic, academic standards, academic of Chinese medicine, academic standards of Chinese medicine in depth, given the logical definition of relevant concepts, and revealed the connotation and logical hierarchy relationship between different concepts. On this basis, we proposed three-level standards points of the academic standards of Chinese medicine. All of these works mentioned above would provide a preliminary standards protection for the academic research of traditional Chinese medicine and the establishment of the modern basic theory of traditional Chinese medicine.
9.Interaction of hypoxia inducible factor expression inhibition and P53 expression of patients with hepatic cellular cancer
Wei ZHANG ; Kun ZHANG ; Zhang DOU ; Tiejun ZHANG
Clinical Medicine of China 2013;29(z1):11-13
Objective To investigate the correlation of hypoxia-inducible factors (HIF),P53 level and liver cancer infiltration in the microenvironment of the liver cancer pathological.Methods Flow cytometry was used to determine the P53 expression.Immunohistochemical method was applied to detect the expression of HIF characteristics,the correlation between Liver function (alanine amiotransferase (ALT),albumin (ALB),alpha fetoprotein(AFP))and HIF,P53.Results HIP positive rate in patients with encapsulated with incomplete,tumor diameter ≤ 5 cm and> 5 cm,tumor stage Ⅰ+Ⅱ and Ⅲ were 50.0% (14/28) and 33.3% (14/42),43.6 % (17/39)and 22.6% (7/31),29.7% (11/37)and 18.2% (6/33)respectively.The HIF positive rate decrease in patients with capsular damage,tumor diameter increases (> 5 cm),Ⅲ patients,and differences between groups were statistically significant (P =0.040,0.030,0.030,respectively).P53 positive expression rate were 42.9% (12/28) vs.83.3% (35/42),51.3% (20/39) vs.93.5% (29/31),35.1% (13/37) vs.69.7% (23/33).The P53 positive expression in patients with capsular damage,tumor diameter increases (> 5 cm),Ⅲ patients with were relatively higher,the difference between groups was statistically significant (P =0.030,0.030,0.040,respectively).HIP positive rate in patient with past suffering from chronic hepatitis,cirrhosis and chronic AFP > 35 ng/L,ALT > 80 U/L were 36.2% (21/58),27.9% (17/61),27.1% (14/51),28.2% (11/39),lower than that of 58.3% (7/12) in non-chronic hepatitis,non-cirrhotic 55.0% (5/9),AFP <35 ng/L 58.0% (11/19),ALT < 80 U/L 51.6% (16/31) of patients,the differences were statistically significant (P =0.046,0.047,0.037,0.045,respectively).P53 positive rate in patient with past sufering from chronic cirrhosis,AFP > 35 ng/L,ALT > 80 U/L,albumin < 35 g/L were 82.0% (50/61),74.5% (38/51),69.2% (27/39),73.7% (42/57),respectively,higher than that of patient without chronic cirrhosis 44.4% (4/9),or with AFP < 35 ng/L 31.6% (6/19),ALT < 80 U/L 22.6% (7/31),albumin > 35 g/L 46.2% (6/13).Tthe differences were statistically significant (P =0.037,0.040,0.032,0.041,respectively).Conclusion HIF and P53 might be the important factor in monitoring the development of hepatocellular carcinoma invasion.
10.Cellular prion protein-mediated hippocampal neuron axon elongation impairment involved in cognitive impairment induced by sleep deprivation
Wei DOU ; Zhen LI ; Zhaohuan ZHANG ; Liuqing HUANG ; Zhongxin ZHAO
Chinese Journal of Neurology 2015;48(1):44-49
Objective To investigate the effect of rapid eye movement (REM) sleep deprivation on spatial memory and hippocampal cellular prion protein (PrPC) expression and to explore the underlying mechanism of cognitive impairment induced by sleep deprivation.Methods Adult Sprague-Dawley rats were sorted by weight,randomly divided into three groups:the cage control (CC) group,the tank control (TC) group,and the sleep deprivation (SD) group.Rats were deprived of REM sleep for 72 h using the modified multiple platform method.The Morris water maze task was used to assess hippocampal-dependent spatial memory.After sleep deprivation,the rats were sacrificed and their brain tissue was analyzed for PrPC protein expression via Western blotting.Hippocampal neuron axon elongation was examined as well after lentivector-mediated RNA interference (RNAi) of PrPC in primary cultured rat hippocampal neurons.Results REM sleep deprivation for 72 h resulted in spatial memory impairment.The number of times of rats passing through the platform was decreased significantly in the SD group (3.17 ±0.95) compared with the CC (7.17 ±0.95) and TC (6.50 ±0.62) groups (Z =2.026 6,Z =2.026 6,P <0.05),the mean value of proximity to the platform (mm) was greater for rats of the SD group (711.74 ± 33.99) compared to those of theCC (592.32±31.31) andTC (580.86±11.36) groups (Z=-2.001 6,Z=-2.4820,P < 0.05).REM sleep deprivation for 72 h resulted in reduced PrPC level in the hippocampus (0.33 ± 0.10) compared with the CC (1.01 ±0.33) and TC (0.96 ±0.27) groups (Z=2.152 9,Z=2.152 9,P < 0.05).In primary cultured hippocampal neurons,axon elongation(μm) was inhibited 7 days in infected neurons (326.28 ± 12.53) compared with normal (555.00 ±30.43) or negative control (558.70 ±23.10) cells (Z =4.768 4,Z =4.877 0,P < 0.05).Conclusion These findings suggest that PrPC-mediated hippocampal neuron axon elongation inhibition is probably involved in spatial memory impairment induced by sleep deprivation in rats.