1.Variation of long-chain 3-hydroxyacyl-CoA dehydrogenase DNA methylation in placenta of different preeclampsia-like mouse models
Yiwei HAN ; Zi YANG ; Xiaoyan DING ; Huan YU ; Yanhong YI
Chinese Journal of Obstetrics and Gynecology 2015;50(10):740-746
Objective By detecting the variation of long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) DNA methylation in preeclampsia-like mouse models generated by different ways, to explore the roles of multifactor and multiple pathways in preeclampsia pathogenesis on molecular basis. Methods Established preeclampsia-like mouse models in different ways and divided into groups as follows: (1) Nw-nitro-L-arginine-methyl ester (L-NAME) group: wild-type pregnant mouse received subcutaneous injection of L-NAME;(2) lipopolysaccharide (LPS) group:wild-type pregnant mouse received intraperitoneal injection of LPS; (3) apolipoprotein C-Ⅲ (ApoC3) group: ApoC3 transgenic pregnant mouse with dysregulated lipid metabolism received subcutaneous injection of L-NAME;(4)β2 glycoprotein I (β-2GPI) group:wild-type pregnant mouse received subcutaneous injection ofβ-2GPI. According to the first injection time (on day 3, 11, 16 respectively), the L-NAME, LPS and ApoC3 groups were further subdivided into:pre-implantation (PI) experimental stage, early gestation (EG) experimental stage, and late gestation (LG) experimental stage.β-2GPI group was only injected before implantation. LCHAD gene methylation levels in placental were detected in different experimental stage. Normal saline control groups were set within wild-type and ApoC3 transgenic pregnant mice simultaneously. Results (1) CG sites in LCHAD DNA:45 CG sites were detected in the range of 728 bp before LCHAD gene transcription start site, the 5, 12, 13, 14, 15, 16, 19, 24, 25, 27, 28, 29, 30, 31, 32, 34, 35, 43 CG sites were complex sites which contained two or more CG sequences, others were single site which contained one CG sequence. The 3, 5, 6, 11, 13, 14, 18, 28 sites in L-NAME, LPS, ApoC3 and β-2GPI groups showed different high levels of methylation; the 16, 25, 31, 42, 44 sites showed different low levels of methylation; other 32 sites were unmethylated. (2) Comparison of LCHAD gene methylation between different groups:the methylation levels of LCAHD gene at 3, 11, 13, 14, 18 sites in L-NAME, LPS, ApoC3 andβ-2GPI groups were significantly higher than those in the normal saline control group (P<0.05); and the methylation levels of 42, 44 sites in these groups were significantly lower than those in the normal saline control group (P<0.05). (3) Methylation of LCHAD gene at the same site between different experimental stages: ① The 3, 11, 18 sites of EG experimental stage was significantly lower than PI and LG experimental stage in L-NAME group (P<0.05);the 3, 11, 18 sites of PI experimental stage was significantly lower than EG and LG experimental stage in LPS group (P<0.05);these sites of PI experimental stage was significantly higher than EG and LG experimental stages in ApoC3 group (P<0.05).②The methylation of site 5 in L-NAME and LPS groups were significantly higher than that of the normal saline control group (P<0.05), and the LG experimental stages were significantly higher than other stages, but in ApoC3 group , only PI and EG stages were significantly higher than the normal saline control group (P<0.05).③At site 6 in L-NAME group which showed high methylation level was significantly higher than the same site in other groups which showed low methylation level (P<0.05).④At 13, 14 sites, earlier preeclampsia onset caused a lower methylation level in L-NAME group, but PI experimental stage was significantly higher than EG and LG experimental stages in LPS group (P<0.05), EG experimental stage was significantly higher than PI and LG experimental stages in ApoC3 group (P<0.05). ⑤ At site 28, earlier preeclampsia onset caused a higher methylation level in L-NAME group, but PI experimental stage was significantly lower than EG and LG experimental stages in LPS group (P<0.05), EG experimental stage was significantly higher than PI and LG experimental stages in ApoC3 group (P<0.05).⑥The 16, 25, 31 sites in ApoC3 group were significantly higher than other groups (P<0.05). ⑦ At site 42 in β-2GPI group was unmethylated, but it in other groups showed low methylation level, the methylation level of site 42 inβ-2GPI group was significantly lower than that in other groups (P<0.05). Conclusions The methylation of 6 and 42 CG sites may be related to LCHAD gene expression in placenta of L-NAME and β-2GPI induced preeclampsia-like models respectively;LCHAD gene expression and DNA methylation may not have obviouscorrelation in LPS and ApoC3 induced preeclampsia-like models. Differences exist in LCHAD DNA methylation in preeclampsia-like models generated by different ways, revealed a molecular basis to expand our understanding of the multi-factorial pathogenesis of preeclampsia.
2.Inhibiting effects of three components of Astragalus membranaceus on oxidative stress in Chang Liver cells.
Jian LI ; Lin HAN ; Yu-fang MA ; Yi-fan HUANG
China Journal of Chinese Materia Medica 2015;40(2):318-323
The main objective of this research is to investigate the effects of astragaloside IV, calycosin separately glucoside, formononetin on oxidative stress in Chang Liver cells induced by H2O2. In the experiments, Chang Liver cells (a kind of normal human hepatocytes) were used as the research object, bifendate which has a clear hepatoprotective effect was used as the positive control drug, then the oxidative damage model of Chang Liver cells were established by H2O2. Cells were divided into six groups: blank control group, oxidative stress group, astragaloside IV group, calycosin separately glucoside group, formononetin group and positive control group. Then endogenous antioxidant system related indexes were detected by micro plate and colorimetric method; intracellular reactive oxygen species (ROS) were detected by DCFH-DA fluorescent probe; and the expressions of CYP2E1 were evaluated by liver microsomes, mRNA, and protein, respectively with spectrophotometry, Real-time PCR method, and Western blot technique. Results showed that H2O2 decreased antioxidant activity, and increased ROS level and expression of CYP2E1. The above oxidative stress status had been changed with protections of the three components of Astragalus membranaceus (compared with oxidative stress group, P < 0.05, P < 0.01), which taken as a whole had equivalent effects as the drug of positive control group( bifendate). Taken together, three Astragalus membranaceus ingredients all had significant or extremely significant inhibiting effects on oxidative damaged Chang Liver cells which were induced by H2O2, and the oxidative damage of Chang Liver cells had been relieved.
Astragalus membranaceus
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chemistry
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Cells, Cultured
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Cytochrome P-450 CYP2E1
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metabolism
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Humans
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Isoflavones
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pharmacology
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Liver
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drug effects
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Oxidative Stress
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drug effects
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Reactive Oxygen Species
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metabolism
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Saponins
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pharmacology
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Triterpenes
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pharmacology
3.The determination of perphenazine by a new flow-injection chemiluminescence method
Jingfeng YI ; Xiaonian HAN ; Chunling YU ; Yuhai TANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2004;0(05):-
Objective To establish a rapid and precise continuous flow-injection chemiluminescence (CL) method for the determination of perphenazine. Methods In HNO_3 medium, perphenazine could be oxidated by ceriuim (IV) and CL was proportional to the perphenazine concentration without any sensitizers. Thus, a new flow-injection CL method was developed. Results Under the optimized conditions, the proposed method allowed the determination range within 1.0?10~ -7 -7.0?10~ -5 g/mL with the detection limit of 8.0?10~ -8 g/mL. Eleven parallel assays were conducted on perphenazine of 1.0?10~ -6 g/mL, with the relative standard deviation of 1.8%. Conclusion The method is simple, rapid, precise, and sensitive, and has broad linear range; therefore, it has been applied to the the determination of the perphenazine in tablets with satisfactory results.
4.Discussion on selection and use of eye lens protective equipment for interventional staff
Weiyue YU ; Faguo CHEN ; Guodong LI ; Yi HAN
Chinese Journal of Radiological Medicine and Protection 2017;37(8):641-644
The decreased occupational dose limit for eye lens has lead to an extensive focus on the eye lens dose monitoring and protection for occupational staff in interventional procedures.Based on the literature investigation of existing measurement and calculation results,the efficiency and influence factors of eye lens protective equipments for interventional staff are analyzed,and the suggestions for selection and use of them are provided.The main contribution to the eye lens dose to interventional staff is unshielded radiation which reaches the eyes directly.The key factors to inflence the efficiency of eye lens protective equipment is the geometric conditions such as structure,beam projection,position arrangement and operator postures,instead of lead equivalent thickness.Equipment of 0.5 mm lead equivalent thickness is enough to protect the eye lens of interventional staff.The combination of lead glasses and lead barrier can provide better protection in clinical practice.
6.Effect of combined therapy of mild hypothermia and hibernation on severe brain injury
Yi-hua AN ; En-zhong LIU ; Chun-jiang YU ; Zhanqiang HAN
Chinese Journal of Rehabilitation Theory and Practice 2004;10(3):181-182
ObjectiveTo investigate the efficacy of combined therapy of mild hypothermia and hibernation to treat severe brain injury. Methods24 patients with severe brain injury were randomly divided into combined therapy group and normothermia group. Glasgow Coma Scale scores of all the patients were in the range of 3 to 8. No later than 10 hours after their injury, hypothermia patients were given half dosage of No.1 hibernation cocktail and had been cooled by cooling blankets to 32℃-34℃ (rectal temperature) for 5 days, then to 35℃ for 24 hours, and slowly increased to their normal level. 3 days and 7 days after their admission, intracranial pressure,creatine phosphate kinase,partial pressure of arterial O2 and CO2, platelet and Na+,K+ were measured.7 days after their admission, Glasgow Outcome Scale scores of each patient and mortality of each group were measured. ResultsThe mortality of combined therapy group(25.0%) was significantly lower than that of normothermia group (66.6%,P<0.05). The decreased values of intracranial pressure, creatine phosphate kinase and platelet number of combined therapy group were all significantly higher than that of normothermia group respectively (P<0.05). There were no significant difference in mean artery pressure, blood electrolyte, and partial pressure of arterial O2 and CO2 between these two groups(P>0.05). ConclusionThe combined therapy of mild hypothermia and hibernation can effectively reduce the mortality of patients with severe brain injury as it is much easier, less invasive and with less complications.
7.Chronic ethanol exposure and withdrawal alter the expression of actin-binding protein and cyclin-dependent kinase-5 in rat brain
Yi YU ; Jinzhu HAN ; Yangyang LIAN ; Jinhong HAN ; Wei HAO ; Ruiling ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2014;23(7):587-590
Objective To investigate the effects of chronic ethanol exposure and withdrawal on the expression of actin-binding protein cofilin,p-cofilin and cyclin-dependent kinase-5 (cdk5) in the nucleus accumbens and striatum in rat brain.Methods Twenty-four male SD rats were randomly divided into one control group and three experimental groups.In the experimental groups,ethanol was administered in drinking water at the concentration of 6% (V/V) for two months.Rats in control group drank normal drinking water.After two months ethanol was removed and ethanol withdrawal syndromes were evaluated.Rats were sacrificed on withdrawal 0 h,withdrawal 6 h and withdrawal 2 d.The expression levels of cofilin,p-cofilin(ser3)and cdk5 in the rat brain were measured by immunohistochemistry methods.Results Withdrawal syndrome scores of ethanol fed rats were obviously higher than those of control rats after ethanol was removed,the highest score occurred at 6 h after ethanol withdrawal.In the nucleus accumbens area of rat brain,the levels of cofilin on withdrawal 0 h significantly decreased compared with control group ((0.31±0.05),(0.39± 0.05),P< 0.05).The levels of cdk5 on withdrawal 0 h and withdrawal 6 h significantly increased compared with control group((0.36±0.07),(0.34±0.07),(0.25±0.05),P<0.05).In the striatum of rat brain,the levels of cofilin on withdrawal 0 h significantly decreased compared with control group ((0.26±0.04),(0.34±0.05),P<0.05).The levels of p-cofilin on withdrawal 6 h significantly increased compared with control group((0.43±0.06),(0.30±0.06),P<0.01).The levels of cdk5 on withdrawal 0 h significantly increased compared with control group((0.35±0.06),(0.26±0.05),P<0.05),and the levels of cofilin on withdrawal 6 h significantly decreased compared with control group((0.37±0.06),(0.26±0.05),P<0.01).Conclusion Chronic ethanol exposure can induce the development of ethanol dependence,and it accompanies with changes in the expression of actin-binding protein and cdk5 in the brain of rats.
8.Influence of prophylactic application of fluconazole on antifungal susceptibility to strains of Candida isolated from the patients with high risk of deep fungal infections
Jin YU ; Min ZHANG ; Ruo-Yu LI ; Wei-Lin XU ; Han-Yun REN ; Yi-Xin SONG ;
Chinese Journal of Laboratory Medicine 2001;0(01):-
8 ?g/ml were 7 strains in prophylactic treatment group and 3 strains in non-fluconazole prophylactic treatment group respectively.The two groups had significant difference (x~2=8.75,P
9.Effects of steaming and baking on content of alkaloids in Aconite Lateralis Radix (Fuzi).
Chang-lin YANG ; Zhi-fang HUANG ; Yi-han ZHANG ; Yu-hong LIU ; Yun-huan LIU ; Yan CHEN ; Jin-hai YI
China Journal of Chinese Materia Medica 2014;39(24):4798-4803
To study the effect of steaming and baking process on contents of alkaloids in Aconite Lateralis Radix (Fuzi), 13 alkaloids were analyzed by UPLC-MS/MS equipped with ESI ion source in MRM mode. In steaming process, the contents of diester-diterpenoid alkaloids decreased rapidly, the contents of monoester-diterpenoid alkaloids firstly increased, reached the peak at 40 min, and then deceased gradually. The contents of aconine alkaloids (mesaconine, aconine and hypaconine) increased all the time during processing, while the contents of fuziline, songorine, karacoline, salsolionl were stable or slightly decreased. In baking process, dynamic variations of alkaloids were different from that in the steaming process. Diester-diterpenoid alkaloids were degraded slightly slower than in steaming process. Monoester-diterpenoid alkaloids, aconine alkaloids and the total alkaloids had been destroyed at different degrees, their contents were significantly lower than the ones in steaming Fuzi at the same processing time. This experiment revealed the dynamic variations of alkaloids in the course of steaming and baking. Two processing methods which can both effectively remove the toxic ingredients and retain the active ingredients are simple and controllable, and are valuable for popularization and application.
Aconitine
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analogs & derivatives
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isolation & purification
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Aconitum
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chemistry
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Alkaloids
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isolation & purification
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Chromatography, High Pressure Liquid
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Drug Stability
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Drugs, Chinese Herbal
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isolation & purification
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Hot Temperature
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Plant Extracts
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isolation & purification
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Steam
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Tandem Mass Spectrometry
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Time Factors
10.Clinical significance of serum VEGF and bFGF detection in patients with non-small cell lung cancer
Ai-qin, GU ; Bao-hui, HAN ; Xue-yan, ZHANG ; Guo-liang, BAO ; Yu, XIN ; Yi-yi, SONG ; Wei-jing, MIAO
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(09):-
Objective To investigate the levels of serum vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) in patients with non-small cell lung cancer(NSCLC) and relationships with c1inicopatho1ogica1 characteristics and their clinical significance. Methods The concentrations of serum VEGF and bFGF were detected by enzyme-linked immunosorbent assay(ELISA) in 40 patients with NSCLC before and after chemotherapy. Results The level of serum VEGF in patients with Ⅳ stage NSCLC was significantly higher than that of Ⅲ stage(P