2.Baicalin Promotes the Neuronal Differentiation of Neural Stem Cells through Downregulating the p-STAT3
Meng CUI ; Shiqing FENG ; Ningjian FAN ; Jun JIA ; Xianhu ZHOU
Tianjin Medical Journal 2013;(8):786-788
Objective To observe the role of baicalin on the expression of phosphorylated protein of signal transduc-ers and activators of transcription signaling proteins (STATs) during the process that neural stem cells (NSCs) differentiating into neurons. Methods NSCs were isolated from the embryonic cerebral cortex of the 14-15-day pregnant SD rats, which were cultured and passaged in vitro. The 3rd generation of NSCs was used in the experiment. NSCs were randomized into nat-ural differentiation control group, three different doses of baicalin groups (7.5μmol/L, 15μmol/L and 30μmol/L), leukemia inhibitory factor (LIF)+basic fibroblast growth factor (bFGF) group and baicalin+LIF+bFGF group. After 6 d culture in vi-tro, the immunohistochemical method was used to observe the expressions of microtubule-associated protein 2(MAP-2) and glial fibrillary acidic protein (GFAP) in different groups. The expression levels of phosphorylation protein of STAT 3 in NSCs were detected by Western blotting method after 2 h and 6 d of culture. Results The expression of MAP-2 in NSCs was in-creased by baicalin, but the expression of GFAP in NSCs decreased. The expression of GFAP in NSCs was enhanced in LIF+bFGF group, which was inhibited by baicalin+LIF+bFGF. The phosphorylation level of STAT3 in NSCs was downregulat-ed by baicalin, but the phosphorylation level of STAT3 was upregulated in LIF+bFGF group. The upregulated phosphoryla-tion level of STAT3 was inhibited in baicalin+LIF+bFGF group(P<0.05). Conclusion Baicalin can induce NSCs to dif-ferentiate into neurons, which may be caused by the downregulation of the phosphorylation level of STAT3 in NSCs.
3.Expression of Caspase-3 mRNA in frontal cortex and hippocampus of chronic stress-induced depression rats treated by electro-acupuncture
Jia LIANG ; Jun LU ; Shanfu CUI ; Junren WANG ; Ya TU
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(2):97-100
ObjectiveTo observe the expression of Caspase-3 mRNA in prefrontal cortex and hippocampus of chronic stress-induced depression rats,and to detect the machnisms of antidepression by electro-acupuncture.MethodsSprague-Dawley rats were randomly divided into four groups:control group,model group,model +electro-acupuncture group and model + paroxetine group,12 rats in each group.Open-field test was used to observe the changes of movements,and real-time fluorescence quantitative PCR (RT-PCR) method was used to detect Caspase-3 mRNA levels in prefrontal cortex and hippocampus.Results ①Open-field test:after stress,compared with control group rats,the model group rats' crossing numbers (29 ± 7),rearing times (6 ± 2) were apparently less than those of control group( (66 ± 13),( 10 ±2) ; P<0.05,P>0.05).In comparison with model group,the crossing times and rearing times being increasing in degree in electro-acupuncture group( (61 ±9),( 13 ±1 ) ) and paroxetine group( (39 ± 10),(8 ± 1 ),P<0.01,P>0.05).② Compared with the control group,Caspase-3 mRNA in prefrontal cortex and hippocampus significantly increased in model group(P <0.05 ) ;and compared with model group,Caspase-3 mRNA in prefrontal cortex in electro-acupuncture group and paroxetine group significantly decreased(P < 0.05 ),and both expression of Caspase-3 mRNA in hippocampus in electro-acupuncture group and expression of Caspase-3 mRNA in hippocampus in paroxetine group decreased (P > 0.05 ).ConclusionChronic stress can increase the expression of Caspase-3 mRNA in prefrontal cortex and hippocampus of chronic stress-induced depression rats,while electro-acupuncture can decrease the expression of caspase-3 mRNA,which may be an important way to anti-depression by electro-acupuncture.
4.Effect of pigment epithelium derived factor on NO and the expression of caspase-3 in retinal tissues of model rats with optic nerve crush injury
Xiao-Xiao, YAN ; Hai-Bo, JIA ; Xiao-Ling, YIN ; Cui, CUI ; Wei-Xing, PU ; Nan, HUO ; Jun-Bo, ZHAO
International Eye Science 2017;17(6):1047-1050
AIM: To analyze the effect of pigment epithelium derived factor (PEDF) on nitrogen monoxide (NO) and expression of cysteine-containing, aspartate-specific proteases-3 (caspase-3) in retinal tissues of model rats with optic nerve crush injury.METHODS: A total of 60 SD rats were randomly divided into the blank control group, model group and PEDF group, with 20 rats in each group.Except the blank control group, the optic nerve crush injury rat models were established in the other groups, and left eyeballs were taken as samples.After successfully modeling, the model group were treated with intravitreal injection of 5μL of balanced salt solution while PEDF group were treated with intravitreal injection of 5μL of PEDF (0.2μg/μL).Two weeks later, the retinal tissues were collected, and changes of shape were observed under microscope after HE staining.The changes of NO level were measured by colorimetry assay, the expression of caspase-3 mRNA and caspase-3 protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot.RESULTS: HE staining showed that retinal tissues of the blank control group arranged neatly and clearly.Retinal ganglion cells (RGCs) arranged in a monolayer, and cells were oval, uniform in size and distribution, the cell nuclei were clear, closely arranged, with clear boundaries.The retinal tissues of the model group were sparse in shape, RGCs showed vacuolar changes, the overall number of cells was reduced, and cell nuclei of residual RGCs showed pyknosis and uneven staining.RGCs in PEDF group were with slightly edema and arranged closely, and the degree of injury was significantly milder than that in the model group.Levels of Caspase-3 mRNA and protein and NO levels in the three groups showed the model group > PEDF group > blank control group (all P < 0.05).CONCLUSION: The application of PEDF can down regulate the expression of Caspase-3 and NO in rates with optic nerve injury and reduce RGCs injury.
5.Relationship of serum prealbumin level with the severity of acute ischemic stroke in 2522 patients
Bin PENG ; Jia SUN ; Jun NI ; Weihai XU ; Lixin ZHOU ; Ming YAO ; Jianming WANG ; Liying CUI
Chinese Journal of Clinical Nutrition 2012;20(1):19-21
Objective To assess the relationship of serum prealbumin level with the severity of acute ischemic stroke (AIS).Methods The clinical data of 2522 AIS patients who were admitted in Peking Union Medical College Hospital from 2000 to 2008 were retrospectively analyzed.The serum prealbumin and hypersensitive C-reactive protein (hs-CRP) levels were evaluated by immunoturbidimetric assay within 24 hours after admission.The AIS severity was evaluated at admission using modified Rankin Scale (mRS).Patients were divided into mild AIS group (mRS score ≤ 3 ) and severe AIS group (mRS score > 3 ).The levels of serum prealbumin and hs-CRP were compared between these two groups,and the relationship between praalbumin and hs-CRP was also analyzed.Results There were 1697 patients in the mild group (average mRS score:1.67) and 825 patients in the severe group ( average mRS score:4.53 ).The severe group had significantly lower prealbumin level [ ( 191.20,82.00)versus (219.18,72.00) mg/L,P =0.000] and higher hs-CRP level [ (6.01,8.22) versus (4.15,6.93) mg/L,P =0.000 ] than the mild group,respectively.Partial regression analysis showed that the stroke severity was negatively correlated with serum prealbumin level (r =-0.115,P =0.000) and positively correlated with hs-CRP level (r =0.098,P =0.000).Serum prealbumin level was inversely correlated with hs-CRP level ( r =- 0.396,P =0.000).Conclusion The prealbumin level is relevant with the severity of AIS and inflammation.
6.Characterization of imipenem non-susceptible Pseudomonas aeruginosa isolates from patients without carbapenem treatment
Yihai GU ; Xiao ZHU ; Shenghui CUI ; Jun ZHANG ; Qingyuan ZHOU ; Jingyun LI ; Jia CHAI
Chinese Journal of Laboratory Medicine 2012;35(8):716-721
Objectives To investigate characterization of imipenem resistance among imipenem non susceptible Pseudomonas aeruginosa isolated from patients who treated without imipenem and explore risk factors of imipenem resistance.Methods From April,2006 to March,2008,a total of 37 non-susceptible to imipenem without imipenem therapy isolates were collected from affiliated 3201st Hospital of Medical College of Xi'an Jiaotong University.The minimum inhibition concentration (MIC) to 11 antimicrobial agents were determined by the broth dilution method.We also tested imipenem MIC combined with efflux pump inhibitor PAβN.PCR was performed to check for the presence of carbapenem-hydrolylzing MBL genes and oprD gene.The expression level of oprD2 and ampC were evaluated by qRT-PCR.Molecular typing was performed using PFGE.Results There is significant difference ( t =- 2.9004,P < 0.01 ) of the average number days of therapy between with two or more antibiotics in the 16 patients (20.0 ± 9.5 ) d and that with only one antibiotic in the other 21 patients ( 12.6 ± 4.4 ) d before imipenem-non-susceptible strains were isolated.In all 37 strains,32 strains showed resistance to more than three antibiotics.The MBL gene ( IMP-9 ) was only found in one strain,but its phenotype is negative,oprD2 gene from the 29 strains were found forward inserted by ISpa1328.Thirty-five isolated were considered to have no oprD expression.The patterns of the total DNA of 37 strains appeared six PFGE types.The 26 strains belonged to C2 PFGE type.In the presence of PAβN,all 37 strains increased sensitivity to meropenem.Conclusion Fluoroquinolones and cephalosporins treatment could play an important role in imipenem non-susceptible production in the research isolates.
7.Risk factors of recurrent ischemic stroke: a retrospective analysis
Bin PENG ; Jia SUN ; Jun NI ; Weihai XU ; Lixin ZHOU ; Ming YAO ; Liying CUI ; Jianming WANG
Chinese Journal of Neurology 2010;43(12):820-823
Objective To examine the recurrent ischemic stroke(RIS)in hospitalized patients predicted by Essen Stroke Risk Score(ESRS)and investigate the effect of intensive antiplatelet therapy in patients with high risk of RIS. Methods Retrospective analysis of patients with acute noncardioembolic ischemic stroke consecutively admitted to Peking Union Medical College Hospital(PUMCH)during 2001-2008. All 1008 patients were included in this study, 377 hospitalized in 2001-2002, 315 in 2005 and 316 in 2008. High risk of RIS was defined as ESRS ≥3. The average ESRS score in three groups and percentage of patients with high risk of RIS in each group were calculated. Risk of RIS was evaluated by comparing the average ESRS score in these three groups. Patients with high risk and taking clopidogrel treatment were also evaluated. Results Average ESRS score was 2. 67 ± 1.47,3.55 ± 1.40 and 3.93 ± 1.48(F= 150. 85,P=0.000)in groups of patients hospitalized in 2001-2002, 2005 and 2008 respectively. A significant difference was shown in percentage of high risk patients with ESRS ≥3 in three groups: as 51.64% in 2001-2002,77. 19% in 2005 and 85. 45% in 2008(X2=98.30,P=0. 000),showing a trend of increasing risk of RIS. Except for patients age, rates of having the risk factors in patients increased significantly, including hypertension, diabetes, myocardial infarction, myocardial diseases, smoking,vascular disease and stroke history. Clopidogrel usage has been increasing to 25.63% in 2008, but was far behind high ratio of high risk RIS patients(85.45%). Conclusions This retrospective study showd an increasing trend in RIS risk during 2001-2008. Identifying high risk patients and applying intensified antiplatelet therapy might play an important role in reducing the risk of RIS.
8.PCR Detection of N.coenophialum and N.lolii
Yue-Ting LIU ; Fang LIAO ; Tie-Jun CUI ; Guo-Ming HUANG ; Jia-Feng LUO ;
Microbiology 1992;0(04):-
18 fungal strains including N.coenophialum,N.lolii, N.huerfanum、N.chisosum、N.aotearoae、N.sp.and 8 varieties of grass seeds belonging to Festuca arundinacea and Lolium perenne have been studied.With amplification of IS1~IS3 and F1~R1 of genomic DNA, the primers Tub-2-F~Tub-2-R from Tubulin-2 gene and F3~R3 from NC25 gene have been designed.A PCR method to detect N.coenophialum and N.lolii was established, and also a nested-PCR method to detect N.coenophialum and N.lolii in single seed was established.These PCR detection methods are strongly special and much credible and rapid-speeded.
9.Clinical observation on electroacupuncture plus Yi Jin Jing (Sinew-transforming Qigong Exercises) for knee osteoarthritis
Jia-Yan DING ; Zi-Yong JU ; Yi-Jun ZHU ; Xu JIANG ; Yue-Hua WANG ; Hua-Shun CUI
Journal of Acupuncture and Tuina Science 2020;18(5):396-402
Objective: To observe the efficacy of electroacupuncture (EA) plus Yi Jin Jing (Sinew-transforming Qigong Exercises) for knee osteoarthritis (KOA). Methods: A total of 60 patients with KOA were divided into an observation group and a control group according to the random number table method, with 30 cases in each group. Patients in the observation group received the treatment of EA plus Yi Jin Jing (Sinew-transforming Qigong Exercises), while patients in the control group only received EA treatment. Both groups were treated for 5 weeks. The changes of Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and visual analog scale (VAS) scores in the two groups were observed after treatment. Results: After treatment, the total effective rate in the observation group (92.3%) was significantly higher than that in the control group (70.0%), (P<0.05); the WOMAC and VAS scores in both groups were significantly lower than those before treatment, showing statistical significance (all P<0.01); there were significant differences in the post-treatment changes in the WOMAC and VAS scores between the two groups (P<0.05, P<0.01). Conclusion: EA plus Yi Jin Jing (Sinew-transforming Qigong Exercises) is clinically effective for KOA. This combined treatment can alleviate clinical symptoms.
10.An experimental study on the reverse mechanism of PPAR-gamma agonist rosiglitazone in rats with non-alcoholic steatohepatitis.
Cai-yan ZHAO ; Ya-dong WANG ; Jun-ying ZHOU ; Bei JIA ; Jun-feng CUI
Chinese Journal of Hepatology 2007;15(6):450-455
OBJECTIVETo investigate the influence and significance of peroxisome proliferator- activated receptor-gamma (PPAR-gamma) agonist rosiglitazone on the expression of I kappa B kinase-beta(IKK-beta) mRNA and protein induced by LPS in Kupffer cells (KCs) cultured in vitro and to investigate the activity of nuclear factor-kappa B (NF-kappa B) together with the expression of cyclooxygenase-2 (COX-2) in livers of rats with non-alcoholic steatohepatitis (NASH).
METHODS(1) KCs from healthy Wistar rats were isolated and purified with IV collagenase digestion and gradient centrifugalization, and then were incubated in the presence or absence of LPS (1 microg/ml) together with two different concentrations of rosiglitazone (10 nmol/L and 50 nmol/L). (2) Thirty-eight healthy Wistar rats were randomly divided into a normal blank control group (10 rats) fed with a normal diet and a NASH model group (28 rats) fed with a fat-rich diet (10% lard + 2% cholesterol + 5% corn oil). After the NASH model was established successfully and confirmed by pathological examination of the livers of 4 rats, 24 rats that continued with the high fat-rich diet, were divided into three groups (8 rats in each group): a control group fed normal saline (NS), a lower dose rosiglitazone group (1 mg.kg(-1).d(-1)) and a higher dose rosiglitazone group (4 mg.kg(-1).d(-1)) for 12 weeks. The mRNA expression of IKK-beta in KCs and COX-2 in livers were measured using reverse transcription-polymerase chain reaction (RT-PCR). The IKK-beta protein in KCs and the NF-kappa B activity of hepatic tissues were determined respectively by Western blot and electrophoretic mobility shift assay (EMSA). The concentration of tumor necrosis factor alpha (TNF alpha) in the supernatant of KCs cultures and serum of the rats was quantified by enzyme linked immunosorbent assay (ELISA).
RESULTSLPS significantly increased the expression of IKK-beta mRNA and protein in the KCs and the concentration of TNF alpha in the supernatant of the KCs cultures. The expressions of COX-2 mRNA and protein were more obvious in rats with NASH than those in the normal control group, and the binding activity of NF-kB correlated positively with the expression of COX-2 in the livers and the level of serum TNF alpha of model rats as well. Rosiglitazone blocked the expression of IKK-beta mRNA and protein induced by LPS in KCs, and also inhibited NF-kappa B activation and reduced COX-2 expression in the rats.
CONCLUSIONSPPAR-gamma specific agonist rosiglitazone can play an anti-inflammatory role by IKK-beta/I kappa B/NF-kappa B/TNF alpha signal ways, and minimize inflammatory reaction at cellular and molecular levels. This may help to provide a new idea for treating NASH effectively.
Animals ; Cyclooxygenase 2 ; metabolism ; Fatty Liver ; drug therapy ; metabolism ; pathology ; Hypoglycemic Agents ; therapeutic use ; I-kappa B Kinase ; metabolism ; Male ; NF-kappa B ; metabolism ; PPAR gamma ; agonists ; Rats ; Rats, Wistar ; Signal Transduction ; Thiazolidinediones ; therapeutic use ; Tumor Necrosis Factor-alpha ; metabolism