1.Cloning and bioinformatics analysis of gene F protein binding protein 1 of hepatitis C virus
Yuan WU ; Yuan HONG ; Jun CHENG
Medical Journal of Chinese People's Liberation Army 1982;0(01):-
Objective To clone and bioinformatically analyze the full-length sequence of F protein binding protein 1 (FBP1). Methods Yeast two-hybrid system was employed to obtain an unknown gene and named it as FBP1.The coding sequence of FBP1 was cloned using molecular biological techniques. Results The coding sequence of FBP1 was cloned successfully. Conclusion FBP1, a cellular protein binding to F protein of HCV, plays an important role in the interaction of virus protein and host cell protein.
2.Hepatocyte growth factor in combination with interleukin-6 to induce the in vitro differentiation of mesenchymal stem cells isolated from human umbilical cord blood into hepatocyte-like cells
Hong YUAN ; Yuzhuo WU ; Hong CHEN
Chinese Journal of Tissue Engineering Research 2007;0(12):-
AIM: Mesenchymal stem cells (MSCs) isolated from human umbilical cord blood can differentiate into hepatocyte-like cells under a suitable induction condition. The optimal condition is still unclear. This study investigated the feasibility of differentiation of MSCs isolated from human umbilical cord blood into hepatocyte-like cells with hepatocyte growth factor (HGF) and interleukin-6 (IL-6) in vitro to find a new cell source for live tissue engineering. METHODS: The experiment was performed from March to September 2007 at the Central Laboratory of First Hospital of Lanzhou University. ①Three cases of umbilical cord blood were collected from full-term pregnant women. Pregnant women had signed an informed consent. The experiment was approved by Hospital’s Ethical Committee. ②MSCs from umbilical cord blood were separated by density gradient centrifugation and adherent method. Cell surface molecule was measured by flow cytometry. Third passage of cells were divided into four groups and cultured in DMEM medium with HGF (HGF group), IL-6 (IL-6 group), HGF+IL-6 (HGF+IL-6 group) or no growth factor (control group). ③The characteristics of proliferation and growth of MSCs were studied by microscopy and methyl thiazolyl tetrazolium (MTT). The phenotypes of MSCs were identified by flow cytometry and immunohistochemical method. Albumin levels in culture supernatants were determined with enzyme-labeled immunosorbent assay (ELISA). RESULTS: ①Growth and division of adherent cells obtained from human umbilical cord blood were good. The shape of MSCs changed into triangle, polygonal or round on days 21-28 in induction groups. ②Positive staining reaction for alpha fetoprotein (AFP) on day 7, for CK18 on days 21, 28 after induction. Albumin production by induced MSCs increased in a time-dependent manner. The positive percents of every hepatocytic marker were higher in the HGF+IL-6 group than in the IL-6 group and HGF group (P
4.Analysis of Chloroform Concentration and the Influence Factors in Public Swimming Pool Water in the Country of Jiading, Shanghai
Hong YUAN ; Qian PENG ; Jie WU
Journal of Environment and Health 1992;0(05):-
Objective To describe the change of chloroform concentration in swimming pool water and to study the influence factors. Methods The swimming pool water samples were collected in 10 public swimming pools and were determined and evaluated according to the Sanitary Standard for Drinking Water(2001), GB/T 17220-1998, GB/T18204.9-2000, GB9667-1997. Results Chloroform concentration in 21 water samples was from lower than 0.006 to 0.184 mg/L, the median was 0.120 mg/L. In the present study, chloroform concentration was correlated positively with turbidity, CODMn, tetrachloride, the free chlorine residual and pH. A negative correlation was found between chloroform concentration and coliform group. Chloroform concentration among the different swimming pools exhibited statistical significance. Conclusion Chloroform concentration in swimming pool water was significantly higher than that in tap water in Jiading, Shanghai. Change of chloroform is different among swimming pools. The increase of chloroform may be related to the pollutants and chlorination in swimming pool water.
5.Preparation and Appraising of Eucalyptus Oil?-cyclodextrin Inclusion Compound
Yuan HE ; Siwang WANG ; Hong WU
China Pharmacy 1991;0(04):-
OBJECTIVE:To prepare and appraise eucalyptus oil?-cyclodextrin inclusion compound,and to testify the feasibility of transforming the dosage form of eucalyptus oil by clathration techniques.METHODS:The physicochemical properties of inclusion compound was identified by thin-layer chromatography(TLC),infrared spectroscopy(IR),ultra-violet spectroscopy(UV)and gas chromatograph mass spectrometer(GC-MS),respectively.Meanwhile,the changes in constituents and clathration outcomes of eucalyptus oil before and after clathration were also investigated.RESULTS:The analytic result of TLC,IR and UV showed that stable inclusion compound has been formed from eucalyptus oil and?-cy?clodextrin.The result of GC-MS demonstrated that there was no significant change in the essential components and the percentage composition of eucalyptus oil before and after inclusion.CONCLUSION:Stable inclusion compound can be made from eucalyptus oil and?-cyclodextrin meanwhile without changes in main components and the percentage composition of eucalyptus oil.
6.Effects of dexmedetomidine on global cerebral ischemia-reperfusion injury in rats
Peipei GUO ; Hong YAN ; Shiying YUAN ; Huisheng WU ; Jingli CHEN
Chinese Journal of Anesthesiology 2011;31(10):1264-1267
Objective To investigate the effects of dexmedetomidine on global cerebral ischemia-reperfusion (I/R) injury in rats.Methods Fifty-four adult male SD rats weighing 200-250 g were randomly divided into 3 groups (n =18 each): shame operation group (group S),global cerebral I/R group (group I/R) and dexmedetomidine group (group D).Global cerebral I/R was produced by occlusion of bilateral common carotid arteries combined with hypotension (MAP maintained at 35-45 mm Hg).In group D dexmedetomidine 3 μg/kg was injected iv immediately after I/R,followed by infusion of dexmedetomidine at a rate of 3 μg· kg- 1 · h- 1 until 2 h of reperfusion.The neurological deficit score (NDS) was assessed (0 =normal,100 =brain death) at 6 h (T1),24 h (T2)and 72 h (T3) of reperfusion.Then six rats were sacrificed in each group and brain tissues were removed for microscopic examination of hippocampus CA1 region and determination of activity of myeloperoxidase (MPO),contents of TNF-α and IL-1β and expression of glial fibrillary acidic protein ( GFAP).Results Compared with group S,NDS,MPO activity and the contents of TNF-α and IL-1β at T1-3 were significantly increased,the expression of GFAP was up-regulated at T2,3 in groups I/R and D ( P < 0.05 or 0.01).Compared with group I/R,NDS,MPO activity and TNF-α concent were significantly decreased at T1-3,IL-1β concent was decreased at T1,2,the expression of GFAP was down-regulated at T2,3 in group D (P < 0.05 or 0.01 ).The pathologic changes were significantly attenuated in group D as compared with group I/R.Conclusion Dexmedetomidine can attenuate global cerebral I/R injury in rats,and the inhibition of inflammatory response may be involved in the mechanism.
7.Effects of glucose and Mg~(2+) in the neurons damaged by glutamate
Hong XING ; Qihua HE ; Lan YUAN ; Jialing XU ; Benji WU
Chinese Journal of Pathophysiology 1989;0(06):-
AIM and METHODS: To observe the effects of glucose-free and Mg 2+ -free in the extracellular fluid on the changes of [Ca 2+ ] i in the cerebro-cortical neurons damaged by 1 mmol/L glutamate using laser confocal scanning microscope. RESULTS: Both frequency and amplitude of neuronal calcium oscillation induced by glutamate were lowered in glucose-free and Mg 2+ -free buffers. The basic [Ca 2+ ] i concentration was lowered in the former case , but it was elevated in the latter case. CONCLUSION: Mg 2+ -free aggravates [Ca 2+ ] i overload induced by 1 mmol/L glutamate ,under certain conditions the glucose-free might resist damage role of glutamate and Mg 2+ -free.
8.Influence of D-galactose on rat spatial learning memory behavior as well as cerebral hippocampal electrophysiology and synaptic morphology
Shujuan YUAN ; Zhixiong ZHANG ; Dingzong WU ; Hong QIU ; Wenlong DING
Chinese Journal of Tissue Engineering Research 2005;9(37):172-175
BACKGROUND: Learning memory disorder is one of the major manifestations of aging. The model of aging induced by D-galactose is a commonly used animal model in recent years, and long-term D-galactose exposure may cause nerve cell morphological changes in animals.OBJETCIVE: To observe spatial learning memory behavior during Dgalactose-induced aging process in order to further explore in vivo evoked long-term potentiation in hippocampus dentate gyrus and synaptic morphological changes in hippocampal CA3 region.DESIGN: Randomized controlled observation.SETTING: Anatomical Teaching and Research Secti , Shanghai Second Medical University; Department of Physiology, Shanghai Traditional Chinese Medicine University.MATERIALS: The experiment was carried out at the Physiological Laboratory of Shanghai Traditional Chinese Medicine University between August 2000 and April 2001. Totally 22 male Wistar rats of 3-month birth age were included and randomized into normal group and D-galactose group with 11 rats in each group. D-galactose was produced by Shanghai No. 2 Chemical Reagent Factory, Morris water maze was home-made by the Institute of Geriatrics, Shanghai Traditional Chinese Medicine university.METHODS: Rats were subjected to hypodermic injection of 1 mL normal saline every day in normal group, or D-galactose of 800 mg/kg daily for 6consecutive weeks in D-galactose group. Rat spatial learning memory behavior was assessed by the latency of Morris water maze; hippocampal dentate gyrus community potentials evoked by monopulse stimulation on perforating fibers were recordedin vivo; meanwhile, the amplitude of monopulse evoked potentials was determined before and after high frequency stimulation, with the amplitude before high frequency stimulation taken as baseline. Transmission electromicroscope was applied in combination with imaging analysis to observe synaptic morphology and structure in rat hippocampal CA3 region. Water labyrinth latency was compared using the variance analysis of repetitive survey design, t-test was used to compare the differences of peak potential latency of community potentials at various time points after long-term potentiation. Moreover, inducing rate of longterm potentiation was compared by χ2-test, XY-540 type biological imaging processing system was used to analyze electromicroscopic pictures, and all available data were analyzed with t-test.MAN OUTCOME MEASURES: [1] Main outcomes: Changes of Morris water maze latency, as well as inducing rate of long-term potentiation and community potentials. [2] Secondary outcomes: Synaptic morphological and structural changes in hippocampal CA3 region.RESULTS: Totally 22 rats were enrolled in this study, with no one lost during water labyrinth test, but one rat in both normal group and D-galactose group died during electrophysiological experiment. Finally 3 rats were randomly selected from each group for electromicroscopic observation. [1]Comparison of the latency for Morris water maze: In contrast with that of normal group, latency for seeking submarine platform was obviously prolonged in D-galactose group [(14.77±10.10), (51.36±12.45) s, P < 0.05].[2] Comparison of evoked potential in hippocampus dentate before high frequency stimulation: The two groups did not obviously differ in community potential amplitude and community potential latency [(1.05±0.47),(0.91±0.41) mV; (5.46±2.09), (5.38±2.26) ms; P > 0.05]. [3] Inducing rate of long-term potentiation in hippocampal dentate gyrus: Compared to that of normal group, inducing rate in D-galactose group obviously reduced after high frequency stimulation (80%, 20%, χ2=7.20, P < 0.01). [4] Comparison of community potential ratio at different time points after high frequency stimulation: Compared to that of normal group, it was notably reduced in D-galactose group at post-stimulation 20, 30, 60 minutes, respectively (1.104±0.196, 0.919±0.162; 1.354±0.212, 0.999±0.219; 1.236±0.174,0.875±0.311; P < 0.05). [5] Comparison of synaptic struc tural parameters in hippocampal CA3 region: Compared to that of normal group, postsynaptic dense bodies became thickened in hippocampal CA3 region of D-galactose group [(40.60±18.26), (26.35±8.15) nm, P < 0.05], the synapse gap increased [(17.69±6.28), (26.95±5.67) nm, P < 0.05] while synaptic active zone was shortened [(265.13±76.50), (229.13±90.68) nm, P < 0.05].CONCLUSION: Hypodermic injection of D-galactose does harm to rat spatial learning memory by reducing the long-term potentiation, inducing rate in rat hippocampal dentate gyrus, attenuating the increase of long-term potentiation-evoked potential amplitude, and even remarkably changing the synaptic ultrastructure in rat hippocampal CA3 region. It suggests that Dgalactose inhibits the long-term potentiation at hippocampal dentate gyrus and affects synaptic structure in hippocampal CA3 region, which is considered as the basis of spatial learning memory behavioral disorder.
9.Protective Effect of Fasudil on Inflammatory Bowel Disease of Rats
Yunling WANG ; Yuan YUE ; Shuangchan WU ; Hong DING
Herald of Medicine 2015;(5):565-569
Objective To investigate the treatment effect of fasudilon inflammatory bowel disease ( IBD) in rats. IBD was induced by using an enema of trinitro-benzene-sulfonic acid ( TNBS ) . Methods A total of 30 female SD rats were randomly divided into normal control group (0. 9% sodium chloride for induction and 0. 9% sodium chloride for treatment), model control group (TNBS for induction and 0. 9% sodium chloride for treatment), and fasudil group (TNBS for induction and fasudil 4. 5 mg·kg-1 for treatment). The disease activity index(DAI) was estimated 14 days later. The MPO activity, TNF-αcontent and IL-1β level in the colon were investigated, while Rho kinase expression was detected by Western blot. The pathological changes were detected by HE staining. Results The DAI of the normal control, model control, and fasudil groups were:(0. 00±0. 00),(7. 76±1. 32),and (3. 20±0. 98), respectively. MPO activity was (59. 32±9. 08),(96. 65±16. 57),and (69. 58±11. 40) U·g-1, respectively. TNF-α was (0. 15±0. 11),(0. 28±0. 22),and (0. 20±0. 62) ng·mL-1, respectively. IL-1β content was (0. 04±0. 01),(0. 08±0. 02),and (0. 06±0. 02) ng·mL-1, respectively. Rho kinase expression was 0. 713± 0. 170,1. 083±0. 210,and 0. 907±0. 260, respectively. Conclusion Fasudil excerts protective effects against IBD in rats by lowering the expression of Rho kinase and attenuating the release of inflammatory mediators, suggesting that Rho kinase could be a new therapeutic target for IBD.
10.Effect of dexmedetomidine on oxidative stress responses during global cerebral ischemia-reperfusion in rats
Peipei GUO ; Huisheng WU ; Hong YAN ; Jingli CHEN ; Shiying YUAN
Chinese Journal of Anesthesiology 2015;35(3):377-379
Objective To evaluate the effects of dexmedetomidine on the oxidative stress responses during global cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty-six male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 3 groups (n =12 each) using a random number table:sham operation group (group S),global cerebral I/R group (group I/R) and dexmedetomidine group (group D).Global cerebral ischemia was induced by occlusion of bilateral common carotid arteries combined with hypotension (MAP maintained at 35-45 mmHg).In group D,dexmedetomidine was infused at a rate of 3 μg · kg-1 · h-1until 2 h of reperfusion after a loading dose of dexmedetomidine 3 μg/kg was intravenously injected immediately after onset of reperfusion.The neurological deficit score (NDS) was assessed at 24 h of reperfusion,the rats were then sacrificed,and their brains were immediately removed for determination of cell apoptosis and levels of malondialdehyde (MDA),superoxide dismutase (SOD) and catalase (CAT).Apoptotic rate was calculated.Results Compared with group S,NDS,apoptotic rate and MDA level were significantly increased,and SOD and CAT levels were decreased in I/R and D groups.Compared with group I/R,NDS,apoptotic rate and MDA level were significantly decreased,and SOD and CAT levels were increased in group D.Conclusion Dexmedetomidine attenuates global cerebral I/R injury through inhibiting the oxidative stress responses.