1.Expression of PED/PEA-15 and XIAP in prostate cancer cells and their effects on prostate cancer cell (PC-3) apoptosis
Xiao-Yong HU ; Xiao-Chun CHEN ; Zhao-Hui ZHU ; Fu-Qing ZENG ; Gong-Cheng LU ;
Chinese Journal of Geriatrics 2001;0(03):-
Objective To study the effect of antiapoptosis factors PED/PEA-15 and XIAP on prostate cancer cells(PC-3)apoptosis.Methods The expressions of XIAP and PED/PEA-15 in prostate cancer cells(PC-3)were respectively assayed using the RT-PCR technique.XIAP and PED/ PEA-15 specific siRNA vectors were designed and constructed and then were transiently cotransfected into PC-3 cells under induction of liposome.The effects of siRNA vectors on PED/PEA-15 and XIAP transcription were assayed by RT-PCR technique,and the effect of XIAP and PED/PEA-15 on cancer cell apoptosis were determined by flow cytometry and microscope observation.Results PED/PEA- 15 and XIAP were both highly expressed in PC-3 cells.Enzyme digestion analysis and DNA sequencing confirmed that the PED/PEA-15 and XIAP-specific siRNA expression vectors were constructed successfully.The designed siRNA sequences of PED/PEA-15 and XIAP could specifically inhibit their transcription.The PC-3 cells which were cotransfected with PED/PEA-15 and XIAP- specific siRNA vectors were more sensitive to doxorubicin.The apoptosis rate of cotransfected cells was significantly increased.Conclusions PED/PEA-15 and XIAP might be involved in the development of prostate cancer.
2.Optimization of Purification Technology for Total Flavonoids from Callicarpa nudiflora Hook. et Arn. by Macroporous Adsorption Resin
Yong LIU ; Pengwei ZHANG ; Wenqin SU ; Xiao LIU ; Keming GONG ; Ni XIANG
Herald of Medicine 2015;(5):640-643
Objective To optimize purification technology of total flavonoids in Callicarpa nudiflora Hook. et Arm. by macroporous adsorption resin. Methods Macroporous resin models including AB-8, D-101, HP-20, HP2MG, were optimized by static adsorption and desorption experiments regarding to adsorption rate and desorption rate of total flavonoids. Purification technology parameters of total flavonoids were optimized by single factor test. Results HP-20 macroporous resin presented the best purification efficiency,the optimum purification conditions were that taking 4. 46 mg·mL-1 of total flavonoidsat pH 3. 0, loading at 3 BV·h-1, washed with 3BV of water at 3 BV·h-1,then eluted with 4 BV 75% ethanol at 2 BV·h-1, finally obtaining the total flavonoids from the dry extract of Callicarpa nudiflora Hook. et Arn. with the purity of 47. 4%. Conclusion HP-20 macroporous resin is suitable for preliminary purification of total flavonoids in Callicarpa nudiflora Hook. et Arn.
3.In vitro lipofectamine mediated NF-κB decoy oligodeoxynucleotides transfection of Kupffer cells
Yong PENG ; Jingdong LI ; Jiangwei XIAO ; Xuhong LI ; Ling GAN ; Jianping GONG
Chinese Journal of General Surgery 2010;25(6):472-475
Objective To study the transfection effects of nuclear factor-KappaB(NF-κB)decoy oligodeoxynucleotides(ODN) to Kupffer cells (KCs) mediated by lipofectamine,and investigate it's suppression effects on KCs activation. Methods Twenty-four Wistar rats were divided into three groups (n=8).(1)Control group,in which the normal KCs were isolated.(2)LPS group,in which 1 ms/L LPs was added to the culture system.(3)NF-κB decoy ODN group,in which KCs were transduced with NF-κB decoy ODN (4μg×105KCs)prior to LPS stimulation.The transfection efficiency Was assayed,and the phagocytosis function,NF-κB(P65) translocation,CD40 mRNA expression of KCs were also detected respectively. Results Kupffer cells were obviously activated after LPS stimulation.the phagocytosis function was reinforced.the activity of NF-κB transloeated from cytoplasm into nucleus was obviosly increaced.The co-stimulatory molecules expression(CD40 mRNA)significantly increased compared with control group(t=4.01,P<0.01).NF-κB decoy oligodeoxynucleotides can efficiently transfected into KCs mediated by lipofectamine,which can obviously suppress KCs activation,and downregulate the expression of downstream gene(compared with LPS group,t=4.89,P<0.01). Condusion NF-κB decoy ODN can efficiently transfect into KCs and inhibit it's activation.
4.Embryo-fetus development toxicity of a novel PPAR-δ agonist in rat.
Hua-Yun GONG ; Yong ZHU ; Zong-He LI ; Xiao-Yan FAN ; Rong FAN ; Fang-Tong WANG
Acta Pharmaceutica Sinica 2014;49(11):1536-1542
The study aims to investigate the embryo-fetus development toxicity of the novel PPAR-δ agonist HS060098 on SD rats. The pregnant rats that were randomly divided into the solvent control group (1% hydroxypropyl methyl cellulose water solution) and HS060098 suspension groups (10, 30 and 100 mg x kg(-1) xd(-1)) were orally administered with HS060098 suspension or vehicle during the gestation of 6 -15 days (GD6-15). At termination (GD20), female rats were sacrificed. The pregnant females were evaluated by corpora lutea count, implantation sites, existence and death of embryos. Fetal sex, weight, externals, variations and malformations of viscus and skeleton were observed. The results show that there were no significant abnormality in maternal general conditions and fetal appearance as well as viscera, but in the 100 mg x kg(-1) x d(-1) group, the maternal weight gain decreased greatly (P < 0.01) and the skeletal ossification delayed remarkably (P < 0.01); in the 30 mg x kg(-1) xd(-1) group, the fatal and litter number of incompletely ossified sternebrae II was higher than those of the control group (P < 0.05); the skeletal malformations occurred in all dose groups, which indicate that the novel PPAR-δ agonist HS060098 had maternal toxicity and adversely effected fetal skeletal development under the experimental conditions.
Animals
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Bone and Bones
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drug effects
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Embryonic Development
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drug effects
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Female
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Fetal Weight
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PPAR delta
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agonists
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Pregnancy
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Rats
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Toxicity Tests
5.Characteristics of fundus autofluorescence and fundus fluorescein angiography in syphilitic posterior uveitis
Yong-hua, LONG ; Wei-jun, WANG ; Yuan-yuan, GONG ; Xiao-dong, SUN
Chinese Journal of Experimental Ophthalmology 2013;(7):621-624
Background The fundus autofluorescence (FAF)can reflect the function of retinal pigment epithelium(RPE) cell.As an invasive examination,it has been extensive used in retina disease,but there has not any report in syphilitic posterior uveitis.Objective This study was to characterize and contrast the FAF and fundus fluorescein angiography (FFA),indocyanine green angiography (ICGA) findings in patients with syphilitic posterior uveitis.Methods A retrospective series of cases observational study was designed.The clinical data of 27 eyes from 18 patients with syphilitic posterior uveitis were included in Shanghai First People's Hospital from 2010 May to 2012 October,and all the patients were diagnosed by serologic and ophthalmic tests.The patients were assigned to acute stage group(with the course <2 months)and chronic stage group(with the course ≥ 2 months).FFA,ICGA and FAF were performed respectively on all the patients,and the examination results were compared and analyzed.Results In the affected eyes with syphilitic posterior uveitis,the FFA image showed a retinal vasculitis sign and mottle-like fluorescence appearance in posterior pole and equator zone,and some affected eyes exhibited edema of optic disc and macula.Hypoautofluorescence zone was seen in the acute stage group and cystoid macular edema was found in the chronic stage group.ICGA presented with a wider damage of RPE,especially in the later phase of ICGA.A confluent of hyperautofluorescence with hypoautofluorescence in the posterior fundus,punctiform hyperautofluorescence as well as hypoautofluorescence in papillitis and macular edema were found on the FAF image.Conclusions The pathological basis of syphilitic posterior uveitis is retinal vasculitis and papillitis.ICGA indicates the damage of choroid membranes and RPE,and FAF reflects a metabolism disorder of RPE in the acute stage and atrophy and loss of RPE in the chronic stage.FAF is helpful for the diagnosis of syphilitic posterior uveitis as an assistant index.
6.The diagnosis and therapy of intractable upper gastrointestinal hemorrhage caused by hepatic arterioportal vein fistulas in hepatocellular carcinoma
Peng-Fei LUO ; Xiao-Ming CHEN ; Li-Gong LU ; Bao-Shan HU ; Yong LI ;
Chinese Journal of Radiology 2001;0(04):-
Objective To investigate the hemodynamics of increasing portal venous pressure(PVP) in hepatocellular carcinoma patients complicated with hepatic arterioportal vein fistulas (HAPVF)and the diagnosis and therapy of intractable upper gastrointestinal hemorrhage caused by HAPVF.Methods One hundred and fifteen cases of hepatocellular carcinoma with upper gastrointestinal hemorrhage were checked by hepatic arteriography and were treated through orifices embolization in cases with severe HAPCF by coils and/or ethanol. Results Twenty-six out of 31 patients suffering intractable upper gastrointestinal hemorrhage have severe HAPVF(the main stem of portal veins are visible).However,there are only 15 patients with light HAPVF among the 84 patients who have mild upper gastrointestinal hemorrhage (the main stem of portal veins are invisible).After the embolization,all of the 26 patients who have severe HAPVF stopped bleeding.Among them,the main stem of hepatic arteries are occluded in 2 patients. Conclusion The existence of severe HAPVF should be taken into consideration when intractable upper gastrointestinal hemorrhage occurs in hepatocellular carcinoma patients,and it can be diagnosed through hepatic artery DSA.Orifice embolization is the most effective method for such kind of hemorrhage.
7.Effects of curcumin on sodium currents of dorsal root ganglion neurons in type 2 diabetic neuropathic pain rats.
Bo MENG ; Lu-lu SHEN ; Xiao-ting SHI ; Yong-sheng GONG ; Xiao-fang FAN ; Jun LI ; Hong CAO
Chinese Journal of Applied Physiology 2015;31(6):541-548
Along with the development of economy and society, type 2 diabetic mellitus (T2DM) has become one of the most common diseases at the global level. As one of the complications of T2DM, diabetic neuropathic pain (DNP) stubbornly and chronically affects the health and life of human beings. In the pain field, dorsal root ganglion (DRG) is generally considered as the first stage of the sensory pathway where the hyperexcitability of injured neurons is associated with different kinds of peripheral neuropathic pains. The abnormal electrophysiology is mainly due to the changed properties of voltage-gated sodium channels (VGSCs) and the increased sodium currents (I(Na)). Curcumin is an active ingredient extracted from turmeric and has been demonstrated to ameliorate T2DM and its various complications including DNP effectively. The present study demonstrates that the I(Na) of small-sized DRG neurons are significantly increased with the abnormal electrophysiological characteristics of VGSCs in type 2 diabetic neuropathic pain rats. And these abnormalities can be ameliorated efficaciously by a period of treatment with curcumin.
Animals
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Curcumin
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pharmacology
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Diabetes Mellitus, Experimental
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complications
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Diabetes Mellitus, Type 2
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complications
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Diabetic Neuropathies
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drug therapy
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Ganglia, Spinal
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cytology
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drug effects
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metabolism
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Neuralgia
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drug therapy
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Neurons
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drug effects
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metabolism
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Rats
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Sodium
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Voltage-Gated Sodium Channels
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physiology
8.Inhibition of Beclin 1 enhances apoptosis by H2O2 in glioma U251 cells.
Xiao-Xia KONG ; Hong-Yu ZHANG ; Zhao-Qin CHEN ; Xiao-Fang FAN ; Yong-Sheng GONG
Acta Physiologica Sinica 2011;63(3):238-244
Oxidative stress could induce apoptosis and autophagy process simultaneously, but the role of autophagy is still not clear. Beclin 1, a key gene regulating the preautophagosome formation, is involved in the injury induced by oxidative stress. To observe the role of autophagy in H2O2-induced injury of U251 cells, the recombinant plasmid Psilencer3.1-siRNA-Beclin 1 was transfected into U251 cells by eukaryotic cell transfection technique. Plasmid vector and cell culture medium were used as negative and control groups respectively. The cells were collected 24 h later, and the cell total protein was extracted to detect Beclin 1, Bcl-2 and Bax protein expressions by Western blot. After the Beclin 1-siRNA cells were treated with 1 mmol/L H2O2, the autophagic vacuoles in the cells were stained with monodansylcadaverine (MDC), and the cell apoptotic ratio was determined with PI/Annexin V-FITC staining by flow cytometry analysis. The results showed that the synthetic siRNA decreased the expression of Beclin 1 protein significantly, but had no obvious effect on the levels of Bcl-2 and Bax protein expressions. Compared with those in the control group, the autophagic vacuoles, the level of LC3-II protein expression and the percentage of apoptotic cells increased (P < 0.05) in 1 mmol/L H2O2 group. In Beclin 1-siRNA + H2O2 group, autophagic vacuoles and the levels of LC3-II protein expression decreased obviously, the percentage of apoptotic cells increased significantly compared with that in 1 mmol/L H2O2 group (P < 0.05). H2O2 and autophagy inhibitor 3-methyladenine (3-MA) combination also increased the percentage of apoptotic cells obviously (P < 0.05). These results revealed that the transfection of Psilencer3.1-siRNA-Beclin 1 effectively inhibited the expression of Beclin 1 protein expression, degraded the autophagy level and increased the apoptotic rate in U251 cells under oxidative stress, which was coincident with the effect of autophagy inhibitor 3-MA. This study suggests that autophagy is a cell protective role in oxidative stress process, and the inhibition of autophagy may enhance apoptosis.
Apoptosis
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drug effects
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Apoptosis Regulatory Proteins
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genetics
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metabolism
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Autophagy
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physiology
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Beclin-1
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Brain Neoplasms
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pathology
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Cell Line, Tumor
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Glioma
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pathology
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Humans
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Hydrogen Peroxide
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pharmacology
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Membrane Proteins
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genetics
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metabolism
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Oxidative Stress
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RNA, Small Interfering
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genetics
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Transfection
9.Role of humoral immune response in the protection induced by H.pylori vaccine with chitosa as adjuvant
Yong XIE ; Yan-Feng GONG ; Nan-Jin ZHOU ; Jiang CHEN ; Xiao-Jiang ZHOU ; Nong-Hua LV ; Chong-Wen WANG
Chinese Journal of Pathophysiology 1986;0(03):-
AIM: To study the immunological protection of H. pylori vaccine with chitosa as adjuvant. METHODS: One-grade female BALB/c mice were randomly divided into nine groups and immunized by ①PBS alone; ②chitosan solution alone; ③chitosan particles alone; ④H. pylori antigen alone; ⑤H. pylori antigen plus chitosan solution; ⑥H. pylori antigen plus chitosan particles; ⑦H. pylori antigen plus CT; ⑧H. pylori antigen plus chitosan solution and CT; ⑨H. pylori antigen plus chitosan particles and CT. At 4 weeks after the last immunization, these mice were challenged by alive H. pylori(1?1012CFU/L) twice at two-day intervals. At 4 weeks after the last challenge, these mice were all killed and gastric mucosa were embedded in paraffin, sectioned and assayed with Giemsa staining. The other gastric mucosa were used to quantitatively culture with H. pylori. ELISA was used to detect H.pylori IgA in saliva and gastric mucosa and anti-H.pylori IgG, IgG1, IgG2a in serum, and immunohistochemical method was used to examine sIgA in gastric mucosa. RESULTS: ①In the groups with chitosan as adjuvant, 60% mice achieved immunological protection, which was according to that with CT as adjuvant (58.33%), and was significantly higher than H. pylori antigen alone and other groups without H. pylori antigen(P0.05)and were significantly higher than those in non-adjuvant groups, while those in the groups with chitosan plus CT were significantly higher than those in the group with CT as an adjuvant(P
10.Effect of puerain on Apelin/APJ system in right ventricle of pulmonary hypertensive rats induced by hypoxia-hypercapnia
Ping HUANG ; Yong-Sheng GONG ; Xiao-Fang FAN ; Qin-Hui ZHANG ; Sun-Zhong MAO ; Liang-Gang HU ;
Chinese Traditional and Herbal Drugs 1994;0(04):-
Objective To investigate whether the effect of puerarin on right ventricle hypertrophy of pulmonary hypertensive rats induced by chronic hypoxia-hypercapnia was related to new peptide Apelin or its receptor(APJ).Methods Thirty male Sprague-Dawley rats were randomly divided into three groups, they are control group,hypoxia-hypercapnia 4-week model group,and hypoxia-hypercapnia 4-week plus puerarin group.The concentrations of Apelin-36 protein in plasma and homogenate of right ventricular muscle were detected by radioimmunoassay.The mRNA expressions of Apelin and APJ in right ventricu- lar muscle were measured by semi-quantitive reverse transcription polymerase chain reaction(RT-PCR). Results The weight ratio of right ventricle to left ventricle plus septum[RV/(LV+S)] in model group was significantly higher than that in control group(P0.05).The plasma concen- tration of Apelin-36 protein in model group was significantly higher than that in control group(P