1.The effect of severe intermittent hypoxia on cognitive function and expression of phosphorylated p38MAPK in rat hippocampus CA1
Hongyang WANG ; Yaning ZHAO ; Baoyuan CHEN ; Xiaoqing HAN ; Panpan ZHANG ; Sa LIU
Chinese Journal of Behavioral Medicine and Brain Science 2012;(11):972-975
Objective To explore the mechanism of severe intermittent hypoxia on oognitive function by evaluatig the effect of chronic intermittent hypoxia on cognitive function,neurons structure,damage,p38MAPK protein expression and neuronal apoptosis in rats hippocampal CA1.Methods Ninety-six mature and male Wistar rats were randomly divided into two groups:control group (UC) and 5% chronic intermittent hypoxia group (5%CIH).Rats in IH groups were suffered 8 hours intermittent hypoxia everyday,and the duration of experiment was respectively 2,4,6 and 8 weeks.After exposed for 2,4,6,and 8 weeks,the cognitive function of rats was assessed with the Morris water maze (MWM) ; the changes in the morphology of nerve cells in hippocampus CA1 region were observed; the expression of phosphorylated p38MAPK protein in hippocampus was detected by the methods of immunohistochemistry and western blot; the apoptosis of nerve cells was detected by the method of TUNEL.Results Compared with control group,with prolonged hypoxia,the time of escape latency obviously prolonged and the time of across the target quadrant shortened significantly in rats of 5% CIH group.The time of escape latency at the 8th week was the longest ((71.71 ± 5.49)s,P< 0.05) in 5% CIH group,and the time of across the target quadrant at the 8th week was the shortest ((26.82 ± 4.30) s,P < 0.05) in 5% CIH group.There appeared neuronal degeneration and necrosis in hippocampus CA1 in 5% CIH group.Compared with the control group,the density of the nerve cells survival in the region of hippocampal CA1 reduced dramatically at the 2nd,4th,6th and 8th week and was the lowest at the 8th week(14.16 ± 2.07,P < 0.05).By Immunohistochemical method,the expression of phosphorylated p38 MAPK of 5% CIH group in hippocampal CA1 was more than UC group at the 2nd,4th,6th and 8th week.By western blot,the expression of phosphorylated p38MAPK of 5% CIH group was more than UC group at the 2nd,4th,6th and 8th week and was the most at the 6th week (2.45 ± 0.14,P< 0.05) ;the index of neuronal apoptosis in hippocampal CA1 was increased significantly at the 2nd,4th,6th and 8th week than UC group and reached to the peak at the 6th week (0.608 ± 0.069,P < 0.05) in the 5 % CIH group.Conclusion Chronic intermittent hypoxia could cause the activation of p38MAPK/pathway of neuronal apoptosis and was important mechanism of cognitive dysfunction at the early and middle stage.
2.The mechanism of quinolone resistance in Pseudomonas aeruginosa
Ke JIN ; Yaping HAN ; Jun LI ; Yinghui LIU ; Yaning MEI ; Yi WEN ; Zuhu HUANG
Chinese Journal of Clinical Infectious Diseases 2009;2(2):74-77
Objective To investigate the mechanism of quinolone resistance in Psendomonas aeruginosa.Methods The minimum inhibitory concentrations (MICs)of ciprofloxacin and levofloxacin with and without carbonylcyainde-m-chlorophenylhydrazone(CCCP)were determined by agar dilution method.Polymerase chain reaction(PCR)and DNA sequencing were used to study the mutations in quinolone resistance-determining region of gyrA and parC genes.The strains were genotyped by enterbacterial repetitive intergenie consensus-PCR(ERIC-PCR).Results Sixteen quinolones-resistant Pseudomonas aeruginosa strains were obtained.The MICs of ciprofloxacin and levofloxacin were not reduced significantly by adding CCCP.Thr-83→Ile of gyrA and Ser-87→Leu of parC were found simultaneously in 16 strains of resistant Pseudomonas aeruginosa.Analysis of ERIC-PCR products indicated that 16 quinolone-resistant strains had an identical band pattern which was different from that seen in the sensitive strains.Conclusion Mutations in gyrA and parC may be the main mechanism of quinolone resistance in clinical isolates of Pseudomonas aeruginosa.
3.CT features of hepatic echinococcosis in children
Xin XU ; Xinyu YUAN ; Xiaoying YANG ; Yaning WANG ; Guihua DONG ; Shujuan HAN ; Haining YU
Chinese Journal of Medical Imaging Technology 2017;33(9):1305-1308
Objective To investigate the CT features of hepatic echinococcosis in children.Methods Clinical and CT data of 27 cases with hepatic echinococcosis confirmed by pathology and/or serology were retrospectively analyzed.Results Totally there were 26 cases of cystic echinococcosis and 1 case of alveolar echinococcosis.Ten cases were with simple hepatic echinococcosis and 17 cases with other organic echinococcosis,18 cases of intrahepatic single hydatid and 9 cases of intrahepatic multiple hydatid.A total of 1 alveolar lesion and 47 cystic lesions were found,including unilocular cyst accounted for 51.06% (24/47),cysts with collapse of the internal capsule accounted for 42.55% (20/47),cysts with wall calcification accounted for 4.26% (2/47) and multiple cysts accounted for 2.13% (1/47).There was no solid lesions.The cyst diameters which were larger than 5 cm accounted for 74.47% (35/47).Conclusion The CT can qualitatively diagnose hepatic echinococcosis in children.It is a reliable method of examination.
4.Expression and significance of CHOP in cognitive dysfunction in rats during intermittent hypoxia
Hongyang WANG ; Lijun DUAN ; Yaning ZHAO ; Guangchao HAN ; Min ZHANG ; Yanan WANG ; Ling WANG ; Jinli CAO
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(3):332-336
Objective To explore the expression changes of C/EBP homologous binding protein (CHOP)in intermittent hypoxic rats with cognitive function decline and clinical significance.Methods We randomly divided 75 adult male Wistar rats into normal group (NC group),5% intermittent hypoxia group (5% CIH group)and 10%intermittent hypoxia group (10% CIH group),with 25 rats in each.Then the three groups were further divided into 3 d,7 d,14 d,21 d and 28 d subgroups,and each time a subgroup had 5 rats.The control group was exposed to the air while 10% CIH group and 5% CIH group were exposed to CIH for eight hours from 8:00 to 1 6:00 each day. After exposure for 3 d,7 d,14 d,21 d and 28 d,the cognitive function of rats was assessed with Morris water maze, the expression of CHOP in the hippocampus was assayed qualitatively by immunohistochemical technique,and the apoptosis of neurons was detected by TUNEL method.Results ① With prolonged hypoxia,the time of escape latency obviously prolonged,the time spent crossing the target quadrant shortened (P < 0.05 ),and the apoptosis index of hippocampal neurons in the CIH groups was increased gradually compared with those in control group (P <0.05).The above-mentioned indexes changed more significantly in 5% CIH group than in 10% CIH group (P <0.05).②.The expression of CHOP protein at each time point was increased (P <0.05 ).In 10% CIH group it reached the peak at 28 d while in 5% CIH group it decreased after it peaked at 21 d.③ The expression of CHOP in the two CIH groups was positively correlated with apoptosis index and animal escape latency time,but negatively correlated with the target quadrant time.Conclusion Intermittent hypoxia,which is likely to induce the high expression of CHOP and activation of neural CHOP mediated apoptosis,causes cognitive impairment of various degrees.
5.Effect of inhibition of P38MAPK phosphorylation on learning and memory and autophagy in hippo-campus of rats after cerebral ischemic postconditioning
Xiaoyun ZHAO ; Ying HAN ; Yaning ZHAO ; Yao LIU ; Jianmin LI ; Na DOU ; Dan LI
Chinese Journal of Behavioral Medicine and Brain Science 2018;27(6):487-491
Objective To investigate the effect of ischemic postconditioning on autophagy and learning and memory impairment in rats with ischemia-reperfusion injury by P38MAPK.Methods Ninty-six rats were randomly divided into sham group ( Sham group), cerebral ischemia reperfusion group ( CIR group),cerebral ischemic postconditioning group ( CIP group) and cerebral ischemic postconditioning com-bined with SB203580 group (CIP + SB203580 group),and 24 rats in each group. The rat model of cerebral ischemia was established by Pulsinelli four-vessel occlusion. The learning and memory abilities of rats were measured by Morris water maze. HE staining were used to detect the morphological changes of hippocampal neurons. The phosphorylation of P38MAPK and Beclin-1,LC3-Ⅱexpression were observed by immunohisto-chemistry. Results Compared with the Sham group,the number of crossing the platform decreased(24 h: (3.04±0.20)times),and the escape latency was longer in CIR group(24 h:(58.38±1.52) s) (all P<0.05). The number of survival neurons reduced (24 h:70.93±1.86),and the expression of P38MAPK,LC3-Ⅱ,Bec-lin-1 in immunohistochemistry were increased in CIR group(all P<0.05). Compared with CIR group,the number of crossing the platform at each time point increased (24 h:(5.46±0.50)times),the escape latency was shorter (24 h:(52.42±1.53)s),the number of survival neurons increased at each time point(24 h:(83.07±5.30)) and the expression level of P38MAPK decreased in the CIP group,while the expression lev-el of LC3-II,Beclin-1 increased (all P<0.05).Compared with the CIP group,the number of crossing the plat-form((24 h:(7.13±0.33)times),the escape latency was shorter (24 h:(48.04±1.39)s),the number of survival neurons increased at each time point(24 h:(91.40±1.74)),and the expression of P38MAPK was down-regulated,while the expression of LC3-II,Beclin-1 were up-regulated in CIP +SB203580 group(all P<0.05). Conclusion Ischemic postconditioning can improve learning and memory impairment in rats with is-chemia-reperfusion injury by P38MAPK regulating autophagy.
6.Protective effect of extracorporeal cardiac shock wave combined with ultrasound microbubble post-conditioning on myocardial ischemia-reperfusion injury in rats
Yajing MIAO ; Yaning XUE ; Xiaoxu WANG ; Hong ZHOU ; Gaojie HAN ; Haijuan HU ; Ruoling HAN ; Hongning YIN
Chinese Journal of Ultrasonography 2024;33(1):77-84
Objective:To investigate the myocardial protective effect of extracorporeal cardiac shock wave therapy (CSWT) combined with sulfur hexafluoride microbubble post-conditioning on myocardial ischemia-reperfusion injury (MI/RI) in rats, and to provide theoretical support for clinical treatment of MI/RI.Methods:A total of 32 male SD rats were randomly divided into 4 groups: sham operation group (Sham group), MI/RI group (IR group), CSWT group (IR+ SW group), and CSWT combined with sulfur hexafluoride microbubble treatment group (IR+ SW+ MB group), with 8 rats in each group. Therapeutic intervention was performed in IR+ SW group and IR+ SW+ MB group on the 1st, 3rd and 5th day after modeling. The left ventricular end-diastolic diameter (LVEDD), left ventricular end-systolic diameter (LVESD), left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) of the rats were measured by echocardiography before and after treatment. On the 7th day, myocardial fibrosis was assessed by Masson staining, and cardiomyocyte apoptosis was observed by TUNEL staining. The myocardial apoptotic proteins Bcl-2, BAX, Cleaved-Caspase-3 and Cleaved-Caspase-9 in the infarct boundary area were detected by Western blot. The differences of the above indexes among different groups were compared.Results:①There was no significant change in heart rhythm and heart rate among the groups before and after treatment, and there was no significant difference in heart rate ( P>0.05). ②The echocardiographic results after treatment showed that, compared with IR group, LVEDD and LVESD in IR+ SW group and IR+ SW+ MB group decreased in turn, while LVEF and LVFS increased in turn with significant differences between each two groups (all P<0.05). ③Compared with IR group, the degrees of myocardial fibrosis and apoptosis in IR+ SW group and IR+ SW+ MB group were alleviated in turn, and the relief in IR+ SW+ MB group was the most obvious. Quantitative analysis showed that compared with IR group, the proportions of cardiomyocyte apoptosis in IR+ SW group and IR+ SW+ MB group decreased in turn, and there were significant differences between each two groups (all P<0.05). ④The results of Western blot detection showed that compared with IR group, the levels of Bcl-2 in IR+ SW group and IR+ SW+ MB group increased in turn, while the levels of BAX and the activation level of Caspase-3 and Caspase-9 protein decreased in turn. These differences were all statistically significant between each two groups (all P<0.05) except for the activation level of Caspase-3 protein between IR+ SW group and IR+ SW+ MB group ( P>0.05). Conclusions:CSWT combined with sulfur hexafluoride microbubble therapy can improve left ventricular remodeling and left ventricular systolic function by inhibiting cardiomyocyte apoptosis.
7.Antimicrobial resistance of gram-negative bacilli isolated from 14 teaching hospitals across China in 2016
Qi WANG ; Ping JI ; Xiuli XU ; Yingchun XU ; Ziyong SUN ; Zhidong HU ; Yunsong YU ; Bin YANG ; Kang LIAO ; Juan LU ; Yunzhuo CHU ; Yaning MEI ; Lizhong HAN ; Xianju FENG ; Hui WANG
Chinese Journal of Laboratory Medicine 2017;40(8):614-622
Objective To investigate antimicrobial resistance among nosocomial gram-negative bacilli in 2016 across China.Methods About 1 394 consecutive and non-repetitive gram-negative bacilli were isolated from 14 teaching hospitals from March to August in 2016 across China.All of these isolates were sent to the central laboratory for reidentification and susceptibility testing.The minimal inhibitory concentration(MICs)of meropenem and other antibacterial agents were determined by agar dilution method.The data were analyzed by using WHONET-5.6 software.Results The activity of antimicrobial agents against Enterobacteriaceae was as follows in descending order of susceptible rate: meropenem (95.2%,891/936), amikacin (94.6%,885/936), ertapenem (92.1%,862/936), piperacillin/tazobactam (88.1%,825/936), imipenem (88.0%,823/936), cefoperazone-sulbactam (83.1%,778/936), cefepime (72.2%,676/936), cefiazidime (72.2%,676/936), levofloxacin(68.8%,644/936), ciprofloxacin (63.2%,592/936), minocyline (62.9%,589/936), cefiriaxone (54.9%,514/936), cefotaxime (54.0%,505/936), cefoxitin (44.3%,415/936).The sensitivities of E.coli to carbapenems, amikacin, piperacillin-tazobactam, polymyxin B and cefoperazone-sulbactam were over 80%.The more sensitive antibiotic to Klebsiella pneumoniae was polymyxin B (99.0%), followed by amikacin (84.9%), meropenem (84.4%) and imipenem (82.0%).The prevalence of extended-spectrum β-lactamase was 62.8%(137/218)in Escherichia coli and 28.3%(58/205)in Klebsiella pneumonia.The activity of antimicrobial agents against E.cloacae, E.aerogenes and Citrobacter freundii was as follows in descending order of susceptible rate: meropenem (97.0%-98.5%), amikacin (95.8%-98.3%), imipenem (94.5%-97.5%), polymyxin B (96.4%-100%), cefoperazone-sulbactam (76.5%-90.0%), ertapenem (73.3%-90.1%), piperacillin/tazobactam (82.4%-88.3%).The most active agents against Pseudomonas aeruginosa were polymyxinB (100%), followed by amikacin (89.3%) and ciprofloxacin (82.4%).The most active agents against Acinetobacter baumannii were polymyxinB (100%).The sensitivities of Acinetobacter baumannii to meropenem, imipenem, minocycline and cefoperazone-sulbactam were 20.3%(39/202), 19.3%(41/202), 66.3%(134/202) and 24.8%(50/202), respectively.Conclusions Carbapenems remain high sensitive against Enterobacteriaceae.Controlling carbapenem resistant Klebsiella pneumoniae is urgent.Drug antimicrobial resistance in A.baumanni is a still serious problem.
8.The Effect of Ischemic Postconditioning on the Notch1 Signaling Pathway and Learning and Memory in Rats with Cerebral Ischemia Reperfusion Injury
Yao LIU ; Xiaoyun ZHAO ; Ying HAN ; Yaning ZHAO ; Jianmin LI ; Jianing LI
Journal of China Medical University 2017;46(11):984-989
Objective To assess the effect of ischemic postconditioning on the Notch1 signaling pathway,and learning and memory in rats with global cerebral ischemia.Methods A total of 128 healthy male SD rats were randomly divided into the sham operation (sham),whole-brain ischemia(CIR),ischemic treatment(CIP),and inhibitor(DAPT) groups,with 32 rats in each group.A global cerebral ischemia model was generated using the modified Pulsinelli's four-vascular occlusion model,and the inhibitor group was injected intraperitoneally with gamma secretase inhibitors (DAPT) before ischemic treatment.Rats' learning and memory function results from the Morris water maze test were assessed by observing the changes in hippocampal CA1 neurons with HE staining.The expression of Notch1 in the CA1 area of the hippocampus was assessed using immunohistochemical and Western blotting methods in rats.Results Compared with the sham group,the hippocampal cell survival rate in the CIR group decreased significantly (P < 0.05) and the expression level of Notch1 increased significantly (P < 0.05).Compared with the CIR group,the CIP group revealed a significantly higher hippocampal cell survival rate (P < 0.05) and an increase in Notch 1 expression (P < 0.05);compared with the CIP group,the hippocampal cell survival rate significantly decreased after DAPT administration (P < 0.05) and Notch1 expression decreased significantly.Conclusion Ischemic postconditioning can promote the recovery of learning and memory ability,which may relate to the activation of the Notch 1 signaling pathway.
9.Synergistic Effects of ERK1/2 and PI3-K Pathway Inhibitors on Autophagy in the Hippocampus of Rats with Subarachnoid Hemorrhage
Chaowang AN ; Yao LIU ; Xiaoyun ZHAO ; Ying HAN ; Yaning ZHAO ; Xu ZHAO ; Jianmin LI ; Chengjing XUE
Journal of China Medical University 2017;46(11):995-1000
Objective To investigate synergistic effects of extracellular signal regulated kinase (ERK1/2) and phosphatidylinositol 3 kinase (PI3-K) pathway inhibitors on autophagy in the hippocampus of rats with subarachnoid hemorrhage (SAH),for identification of therapeutic targets in SAH.Methods Totally,200 male SD rats were randomly divided into a sham operated group,SAH group,inhibitor U0126 group,inhibitor LY294002 group,and a U0126+ LY294002 group.Animal models were established by injecting autologous blood twice into the cisterna magna.Morphological changes in the hippocampus nerve cells were detected by HE staining;ERK1/2,PI3-K,beclin-1,and LC3 mRNA expression in the hippocampus were detected by real-time PCR,and phosphorylated ERK 1/2,PI3-K,beclin-1,and LC3-Ⅱ protein expression were detected by immunohistochemistry.Results Neuronal death rate and phosphorylated ERK1/2,PI3-K,beclin-1,and LC3-Ⅱ levels in the hippocampus in the SAH group were higher than in the sham group (all P < 0.05).Neuronal death rate in U0126 or LY294002 group was higher than in SAH group,while ERK1/2,PI3-K,beclin-1,and LC3 mRNA and phosphorylated ERK 1/2,PI3-K,beclin-1,and LC3-Ⅱ protein levels were lower than in SAH group (all P < 0.05).Neuronal death rate in U0126 +LY294002 group was higher than in U0126 or LY294002 group,while ERK1/2,PI3-K,beclin-1,LC3 mRNA and phosphorylated ERK 1/2,PI3-K,beclin-1,and LC3-Ⅱ protein levels in the hippocampus were lower than in U0126 or LY294002 group (all P < 0.05).Conclusion Co-targeting the inhibition of ERK 1/2 and PI3-K pathways can significantly reduce neuronal cell autophagy and aggravate cells loss after SAH.
10.Effect of Plantaginis semen on the expressions of AQP4 gene and protein in small intestine of diarrhea rats
Yaning BIAO ; Ruilong WANG ; Xinqiao CHU ; Weiwei HU ; Nabo ZHANG ; Xue HAN ; Yixin ZHANG ; Chen LIU
International Journal of Traditional Chinese Medicine 2020;42(8):758-762
Objective:To explore the effect of Plantaginis semen on the expressions of AQP4 gene and protein in small intestine of diarrhea rats. Methods:Sixty Sprague-Dawley male rats were randomly divided into normal control group, model control group, hydrochlorothiazide group, and low-, medium-, and high-dose Plantaginis semen groups. Intragastric administration of Senna (20 ml/kg) was implemented in the morning for 5 groups except the normal group. The rats in the low-, medium-, and high-dose Plantaginis semen groups were intragastrically administered with 0.95, 1.9, 3.8 g/kg Plantaginis semen formula granule solution, while the rats in hydrochlorothiazide group were given hydrochlorothiazide suspension 9 mg/kg by gavage according to 1 ml/100 g body weight for 14 days. The loose stool rate, average ranking of loose stool, and diarrhea index were compared according to the fecal traits and stool times after 14 days of treatment, and the small intesine tissue were collected. Hematoxylin-eosin (HE) staining to observe the pathological morphological changes in small intestine, and Quantiative Real-time PCR and Western blot were applied to detect the gene and protein expressions of AQP4. Results:Compared with the model group, the loose stool rate (50.89% ± 6.17%, 41.14% ± 4.48%, 36.37% ± 4.81 % vs. 67.45% ± 7.35%), the average ranking of loose stool (2.16 ± 0.34, 1.73 ± 0.28, 1.52 ± 0.25 vs. 2.63 ± 0.29), and the diarrhea index (1.10 ± 0.19, 0.71 ± 0.11, 0.57 ± 0.12 vs. 1.77 ± 0.24) of rats in each group of Plantaginis semen significantly decreased ( P<0.05); the degree of intestinal mucosal injury, hyperemia and neutrophil infiltration were alleviated; the expressions of AQP4 mRNA (0.48 ± 0.10, 0.69 ± 0.12, 0.97 ± 0.15 vs. 0.21 ± 0.03), and the protein of AQP4 (0.59 ± 0.08, 0.64 ± 0.09, 0.78 ± 0.11 vs. 0.32 ± 0.05) in the small intestine tissue of Plantaginis semen groups significantly increased ( P<0.05). Conclusions:Plantaginis semen has antidiarrheal effect, and its mechanism is related to up-regulation the gene and protein expressions of AQP4, addition of water absorption and promotion of water and fluid metabolism.