1.Interaction between methylenetetrahydrofolate reductase C677T gene polymorphism and sleep duration on risk of stroke pathogenesis
Yan ZHANG ; Ruping XIE ; Yang SHEN ; Dongsheng FAN
Journal of Peking University(Health Sciences) 2008;40(3):262-269
Objective: To investigate the interaction between methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism and sleeping duration on risk of stroke in a Chinese Han ethnic population. Methods: In the case-control study and epidemiological investigation, the self-reported sleep duration and MTHFR C677T polymorphism of 245 patients with cerebral infarction, 222 patients with cerebral hemorrhage and 282 controls were collected. Multiple logistic regression was performed to analyze the interaction between MTHFR C677T polymorphism and sleeping duration on risk of stroke. Results: After adjustment for major confounding variables, multiple logistic regression analysis showed that: (1) There was significant association between long sleep duration (>8 hours of sleep per night) and cerebral infarction (OR=3.90; 95% CI:2.43-6.26), but not for cerebral hemorrhage (OR=1.16, 95% CI: 0.71-1.92). On the other hand, insomnia (sleep duration less than 6 hours) was neither associated with cerebral infarction nor hemorrhage. (2) MTHFR TT genotype increased the risk of cerebral infarction significantly (OR=2.01; 95% CI:1.12-3.61), but not for cerebral hemorrhage (OR=1.16,95% CI:0.71-1.92). (3) There was a significant synergistic effect of interaction between MTHFR TT677 genotype and long sleep duration on risk of cerebral infarction (OR=6.22; 95% CI:2.44-15.83). Conclusion: MTHFR TT677 genotype and long sleep duration increase the risk of cerebral infarction independent of confounding factors, respectively. Furthermore, there is a significant synergistic effect between MTHFR TT677 genotype and long sleep duration on risk of cerebral infarction in the Chinese Han ethnic population.
2.Construction of a cDNA Library and Cloning of an Arabinosidase cDNA from Armillariella tabescens
Dongsheng YAO ; Hui HUANG ; Long ZHAO ; Chunfang XIE ; Daling LIU
China Biotechnology 2005;25(6):65-70
The expression cDNA library of A. tabescens was constructed by SMART technique, which useλTriplEx2 as a vector. The titer and the percentage of the constructed library were about 1.0 × 106pfu/mland 98.3% respectively, and the titer and the capacity of the amplified library were about 3.1 × 108pfu/mland 4.2 × 1010. The library was used to provide expressed sequence tags (ESTs). 147 Expressed SequenceTaqs (ESTs) were gained from 176 clones, which were selected randomly and sequenced at the 5'end. Thesequences were submitted to the EMBL database. Blasting the sequences in the GenBank, 43 of them werefound that they have significant similarity with data in GenBank. EST AJ620046 was has significantsimilarity with the arabinosidase of Bacteroides thetaiotaomicron. Using SMART-RACE a full-length cDNA ofAJ620046 was successfully obtained. In order to initially characterize the biochemical properties ofAJ620046, the ORF of AJ620046 named AF was cloned and expressed in Pichia Pastoris yeast.Recombinant pHIL-S1-AF constructed by inserting AF into pHIL-S1 was transformed into Pichia PastorisGS115. Preliminary experiments indicated that AJ620046 was expressed as a 32 kDa protein in recombinantyeast.
3.Reparation of large coloboma raw surface after facial tumour resection in elderly patients
Honghong LI ; Dongsheng CAO ; Juan XIE ; Qiong BAO ; Yin LOU
Chinese Journal of Geriatrics 2016;35(1):68-71
Objective To investigate the surgical reparation of large coloboma raw surface after facial tumour resection in elderly patients.Methods According to the position and characteristics of tumor as well as the age and tolerance of the patients, full thick skin graft, the skin flaps with subcutaneous pedicle and free skin flap were designed and used in the reparation.Results 24 cases with large coloboma raw surface (5 cm×7 cm-12 cm× 16 cm)were treated by the utilization of three approaches after tumor resection.The large coloboma raw surface in all patients achieved the healing with satisfactory appearance.Conclusions After facial tumour resection, the large coloboma raw surface can be repaired by using the skin graft, skin flaps after tumor resection or free skin flap if designed reasonably.The procedure of operation is simple and the therapeutic effect is satisfactory.
4.Effect of salinity and temperature on motility of Vibrio parahaemolyticus
Xinbo DONG ; Yiquan ZHANG ; Ruifu YANG ; Chuanxiao XIE ; Dongsheng ZHOU
Military Medical Sciences 2014;(12):962-964
Objective To investigate the effect of salinity and temperature on motility of Vibrio parahaemolyticus. Methods V.parahaemolyticus was inoculated on swarming or swimming agar plates containing different amounts of salinity (0.5%, 1.0%, 2.0%, and 4.0% NaCl, respectively), followed by incubation at 26 or 37℃, before the diameters of bacterial lawns were measured .Results and Conclusion The swarming motility was not affected by salinity , while the swimming motility was positively correlated with salinity .Maximum swimming occurred in 2.0% NaCl, and displayed a slight decline in salinity of 4.0%.Both swimming and swarming were affected by temperature , and the motility was signifi-cantly enhanced in 37℃vs 26℃.These results indicate that both salinity and temperature can modulate the motility of V. parahaemolyticus.
5.Effects of NS-398 on the proliferation and apoptosis of HepG2 cells
Haiyang XIE ; Xiao XU ; Shusen ZHENG ; Tingbo LIANG ; Dongsheng HUANG
Chinese Journal of Pathophysiology 2000;0(07):-
AIM: To evaluate the effects of NS-398, a cyclooxygenase-2(COX-2) inhibitor, on the proliferation and apoptosis in HepG2 cells. METHODS: The effects of NS-398 on the proliferation of HepG2 cells was evaluated by MTT. DNA fragmentation gel analysis was used to analyze the apoptotic cells; DNA ploidy and apoptotic cell percentage were examined by flow cytometry. Furthermore, the expression of COX-2 and Bcl-2 mRNA was identified by competitive RT-PCR. RESULTS: NS-398 inhibited cell proliferation and induced apoptosis in HepG2 in a concentration-dependent manner. DNA ploidy analysis showed that S phase cells were significantly decreased with NS-398 concentration increasing. The quiescent G_0/G_1 phase was accumulated with decreasing of Bcl-2 mRNA. Whereas NS-398 had no effect on the expression of COX-2 mRNA, no correlations were found between COX-2 mRNA and the HepG2 cell proliferation and apoptosis induced by NS-398 (r=0.056 and r=0.119, respectively). CONCLUSION: NS-398 significantly inhibits the proliferation and induces apoptosis in HepG2. Mechanisms may be involved in accumulation of quiescent G_0/G_1 phase and decrease in Bcl-2 mRNA expression, but independent to COX-2 mRNA expression. [
6.Study on Improving the Teaching Quality in Respiratory Department Training
Bin HUANG ; Dongsheng LU ; Yongping ZHENG ; Zhixiong CAI ; Canmao XIE
Chinese Journal of Medical Education Research 2003;0(04):-
Some measures about improving the teaching quality in respiratory department training are described in this report,including identifying the aim and importance of major training,raising students'enthusiasm,enhancing the basic major procedure training,performing various teaching,concentrating on the students'response and altering teaching methods continuously
7.Effect of LX A4 and BML-111 on TLR4/NF-κB signaling pathway in macrophage
Daze XIE ; Lixing HUANG ; Dongsheng LIU ; Jun ZHU ; Yong XIE ; Nanjin ZHOU
The Journal of Practical Medicine 2015;(17):2799-2802
Objective To determine the effect of lipoxins (LX) A4 and its agonist (BML-111) on the survival of RAW264.7 macrophage cells and TLR4/NF-κB signaling pathway. Methods RAW264.7 cells were treated with different concentrations of LPS, then the effect of LX A4 and BML-111 on the survival rate of these cells was observed. Cytotoxicity were detected by CCK-8 method and RT-PCR was used to detect the TLR4 and TRAF6 mRNA. The protein levels of TLR4 and pNF-κB p65 in RAW264.7 were determined by Western Blot. Results The survival rates of macrophage treated with LPS for 6 h in 1 000 ng/mL LPS in LX A4 group and BML-111 group were significantly higher than that in control group (P < 0.05). In the present of LPS, the TLR4 mRNA levels in RAW264.7 cells from LX A4 group and BML-111 group were significantly higher than those in the corresponding non-LPS groups. And the TRAF6 mRNA levels in each LPS stimulation group were higher than those in the corresponding non-LPS groups (P<0.05), while the protein level of TRAF6 in LX A4 and BML-111 groups were significantly lower than that in control group (P < 0.05). Stimulated with LPS, the protein levels of pNF-κB p65 in the LX A4 group and BML-111 group were all significantly lower than that in control group (P<0.05), and pNF-κB p65 expression level in control group was also significantly higher than the corresponding non-LPS groups (P < 0.05). Meanwhile, no significant difference was found between LX A4 and BML-111 group (P > 0.05). Conclusion LX A4 and BML-111 could inhibit the cytotoxicity of LPS on the RAW264.7 macrophage cells through the inhibition of the activation of TLR4/NF-κB signaling pathway, then reduce the inflammation. And this stable BML-111 may appear as another promising treatment for IBD disease.
8.Antigrowth effects of chitosan and its derivatives on human hepatocellular carcinoma cell line SMMC7721
Yong XIE ; Nanjin ZHOU ; Jun CAO ; Bin DING ; Dongsheng LIU ; Jinlin LIU
Chinese Journal of Tissue Engineering Research 2008;12(23):4579-4582
BACKGROUND: Different derivatives of chitosan with different molecular weights or degrees of deacetylation show different anti-tumor effects.OBJECTIVE: To study the inhibition effect of water-soluble chitosan and its derivatives, such as sulfonated chitosan, carboxymethyl chitosan and chitooligosaccharides for the growth of hepatocellular carcinoma cell line SMMC7721.DESIGN, TIME AND SETTING: Controlled experiments based on observation were carried out in Jiangxi Institute of Digestive Disease (Nanchang, Jiangxi, China) from January 2004 to December 2006.MATERIALS: Hepatoma cell line SMMC7721 was provided by Jiangxi Institute of Digestive Disease (China). 85.5% deacetylated chitooligosaccharides and 85% deacetylated water-soluble chitosan were produced by Jinan Haidebei Ocean Biological Engineering Co., Ltd (China); Carboxymethyl chitosan and 88.5% deacetylated chitosan were the products of Shanghai Qisheng Biological Products Co., Ltd (China).METHODS: Sulfonated chitosan was prepared using 88.5% deacetylated chitosan and chlorosulfonic acid-formamide, and then was detected with infrared spectroscopy in the Detection Analysis and Test Center, East China University of Science and Technology. SMMC7721 cells in the log phase were inoculated into 96-well culture plates, which were then added with water-soluble chitosan, sulfonated chitosan, carboxymethyl chitosan and chitooligosaccharides with the final concentrations of 25, 50, 100, 200, 400 and 800mg/L. This test was repeated for 3 times, while the control group was also set each time. After 72 hours of routine culture, MTT solution was added into each well and inoculated for another 4 hours. After the culture was terminated, dimethyl sulfoxid was added. The absorbance value of each well was measured at 490nm wavelength on a microplate reader. Three tests were measured to obtain the mean value. Also the inhibition rate was calculated.MAIN OUTCOME MEASURES: Growth inhibition effect of chitosan and its derivatives on the hepatoma cell line SMMC7721.RESULTS: Among the chitosan and its derivates at four kinds of concentrations, water-soluble chitosan and sulfonated chitosan could significantly inhibit the growth of SMMC7721 cells (P<0.001), and the effect was the most significant in the case of sulfonated chitosan. Treatment with water-soluble chitosan and sulfonated chitosan at the concentration of 50mg/L could inhibit the growth of SMMC7721 cells in a dose-dependent manner, and reached a peak at the concentration of 400mg/L and 800mg/L, respectively. Carboxymethyl chitosan and chitooligosaccharides showed no growth inhibition effect (P>0.05).CONCLUSION: Water-soluble chitosan and sulfonated chitosan have significant antigrowth effects on hepatoma carcinoma cells, while carboxymethyl chitosan and chitooligosaccharides are ineffective.
9.The dynamic changes of plasma yon willebrand factor and P-selectin in the finger replanted patients and their clinical significance
Lanfen PENG ; Dongsheng CHEN ; Wenjin FU ; Jincai LUO ; Guangzhong XIE ; Changqin YE ; Zhihong HUANG ; Huihua TANG
Chinese Journal of Laboratory Medicine 2008;31(10):1157-1160
Objective To explore the dynamic changes of von willebrand factor(VWF)and Pseleetin in the finger-replanted patients,and the relationship between the prognosis of the surgery and hypercoagulability.Methods From December 2004 to December 2006,eishty finger-replanted patients were recruited to our study.with 40 healthy volunteers as controls.Plasma VWF and P-selectin were detected by enzyme-linked immunosorbent assay(EUSA)in both controls and patients before or after replantation.Results The VWF and P-selectin levels had significant differences between the replantations and controls(F=14.76,11.76,P<0.01).The VWF levels in the patients of 1,4,8,16 hours after replantation were(1 715±493),(1 396±549),(1 266±504),(1 163±436)U/L respectively,all markedly higher than the controls(P<0.01).The P-selectin levels in patients of 1,4,8,16,24 hours after operation were(14.7±2.6),(12.5±3.0),(11.8±3.2),(11.1±3.0)、(10.5±2.6)μg/L,significanfly higher than the controls(P<0.01).The VWF levels in patients of pre-replantion and the 1,4,8,16,24,48,72 hours after replantation were(854±209),(1 535±389),(1 177±407),(1 040±283),(958±216),(829±193),(777±151),(713±137)U/L in successful group,and were(1 202±164),(2 333±243),(2 146±161),(2 039±244),(1 865±170),(1 645±283),(1 427±331),(1 188±262)U/L in unsuccessful groups.They were all significantly different at the same test-time points between two groups(t=4.44,5.12,6.10,8.43,10.17,8.85,5.10.4.61,P<0.05).The P-selectin levels in patients of 1,4,8,16,24,48,72 hours after replantation were(13.9±2.5),(11.2±2.0),(10.2±1.6),(9.6±1.2),(9.2±0.9),(9.5±0.6),(9.3±0.4)μg/L in successful group,and(17.2±1.0),(16.9±1.0),(17.0±1.3),(16.1±1.1),(14.9±1.5),(13.8±1.4),(12.8±1.2)μg/L in unsuccessful group.Significant difference existed at the same testtime points between two groups again(t=5.22.9.91,10.35,12.79,9.46.9.45,9.33,P<0.01).After replantation,both VWF and P-selectin were rapidly elevated and went to the summit 4 hours later,then declined to pre-replantation level about 24 to 48 hours later after replantation.Conclusions VWF and P-selectin were associated with the hypercoagulability.Dynamic monitoring VWF and p-selectin may be useful in determining the existence of hypercoagulability and the therapy of anti-coagulability.
10.A Preliminary Study of Laryngeal Electromyography for Evaluating Swallowing Function of Patients with Amyotrophic Lateral Sclerosis
Huashun XIE ; Furong MA ; Dongsheng FAN ; Liping WANG ; Yan YAN ; Peiquan LU
Journal of Audiology and Speech Pathology 2014;(5):485-488
Objective To investigate the clinical values of laryngeal electromyography (LEMG) for evaluating the amyotrophic lateral sclerosis(ALS) patients with swallowing disorder .Methods A total of 25 ALS patients with or without complaining of swallowing disorder were recruited for the study .Their right cricothyroid muscles were taken unilaterally with concentric needle electrodes by anterior cervical approach .Each patient was tested with Kub-ota drinking test too .Results Of all the 25 patients ,11 were found abnormal in the right cricothyroid muscle which presented with typical degeneration patterns ,15 cases were found abnormal by Kubota drinking test .The coinci-dence rate of the two methods was 60% ,the poisitive coincidence rate was 32% .The McNemar test showed there was some consistency between the drinking water test and LEMG (P=0 .344) ,and the Kappa coefficient was 0 .219 . Conclusion The cricothyroid muscle electromyography may also be a potential method to evaluate ALS patients since it seemed to offer an evidence of dysphagia related with abnormal sensory function of laryngeal mucosa .But there may be some limitations .