Mechanism of microRNA-223-3p inhibiting hepatic stellate cell activation by targeting microtubule-associated protein 1B / 临床肝胆病杂志

ObjectiveTo investigate the effect of microRNA-223-3p （miR-223-3p） on hepatic stellate cell （HSC） activation and its mechanism. MethodsHuman HSC LX2 cells were selected for the study， and LX2 cells were stimulated by TGF-β to establish a model of HSC activation； quantitative real-time PCR was used to measure the change in the expression level of miR-223-3p during HSC activation. After LX2 cells were transfected with miR-223-3p mimic， quantitative real-time PCR， Western blot， and immunofluorescence assay were used to clarify the regulatory effect of miR-223-3p on HSC activation， and dual-luciferase reporter assay was used to verify the association between miR-223-3p and the target gene MAP1B. After LX2 cells were transfected with MAP1B siRNA， Western blot was used to clarify the influence of inhibiting MAP1B expression on HSC activation； after LX2 cells were transfected with miR-223-3p， quantitative real-time PCR and Western blot were used to verify the regulatory effect of miR-223-3p on MAP1B. The independent-samples t test was used for comparison of continuous data between two groups. ResultsHSC in the activated state had a significant reduction in the expression level of miR-223-3p compared with those in the resting state （t=9.12， P<0.001）. Overexpression of miR-223-3p inhibited the mRNA and protein expression levels of the markers for HSC activation alpha-smooth muscle actin and collagen type Ⅰ （mRNA expression： t=8.35 and 12.23， both P<0.01； protein expression： t=16.24 and 20.90， both P<0.001）. The dual-luciferase reporter assay confirmed that MAP1B was a potential target gene of miR-223-3p. Compared with the control group， LX2 cells with miR-223-3p overexpression had significant reductions in the mRNA and protein expression levels of MAP1B （mRNA expression： t=5.95， P<0.01； protein expression： t=11.12， P<0.001）. ConclusionThis study shows that miR-223-3p can inhibit HSC activation by targeting MAP1B.

Effect of Aspergillus fumigatus on the autophagic flux in murine macrophages: a preliminary study / 中华皮肤科杂志

Objective:To explore the effect of Aspergillus fumigatus ( A. fumigatus) on the autophagic flux in murine bone marrow-derived macrophages (BMDM) . Methods:Murine BMDM were in vitro cultured with heat-killed A. fumigatus for 0, 0.5, 4, and 12 hours. Then, cellular proteins were extracted, and Western blot analysis was performed to detect the conversion of the key autophagy protein microtubule-associated protein 1 light chain 3 (LC3) -Ⅰ to LC3-Ⅱ, and to determine the protein expression of phosphorylated mammalian target of rapamycin (p-mTOR) Ser2481. Additionally, murine BMDM were in vitro cultured with A. fumigatus alone or in combination with different lysosomal inhibitors, including the cysteine cathepsin inhibitor E-64d + pepstatin, bafilomycin-A1 (BAF-A1) , ammonium chloride (NH 4Cl) , and chloroquine, for 4 or 12 hours. Then, Western blot analysis was performed to investigate the effect of A. fumigatus on newly formed LC3-Ⅱ and basal autophagic flux, and confocal laser scanning fluorescence microscopy to analyze the colocalization of A. fumigatus with LC3 and Rubicon (a RUN domain Beclin-1-interacting and cysteine-rich-domain-containing protein) . Experimental results at different treatment time points were analyzed by using unpaired t test, and results of experiments evaluating the effect of two factors ( A. fumigatus spores and autophagosome inhibitors) were analyzed by 2 × 2 factorial analysis. Results:After in vitro co-culture with A. fumigatus for 0.5, 4, 12 hours, Western blot analysis showed that the conversion of LC3-Ⅰ to LC3-Ⅱ increased over time in murine BMDM compared with the control (0 hour) group ( t = 6.58, 3.28, 3.02, respectively, all P < 0.05) , but the protein expression level of p-mTOR (Ser2481) did not significantly differ at different treatment time points ( t = 0.441, 0.477, 0.382, P = 0.682, 0.660, 0.722, respectively) . After 4- and 12-hour in vitro treatment, the accumulation levels of LC3-Ⅱ in BMDM significantly increased in the A. fumigatus + chloroquine group compared with the chloroquine-alone group ( t = 2.13, 2.78, respectively, both P < 0.05) , in the A. fumigatus + NH 4Cl group compared with the NH 4Cl-alone group ( t = 2.92, 2.92, respectively, both P < 0.05) , in the A. fumigatus + BAF-A1 group compared with the BAF-A1-alone group ( t = 2.13, 2.13, respectively, both P < 0.05) , and in the A. fumigatus + E-64d + pepstatin group compared with the E-64d + pepstatin group ( t = 2.13, 2.92, respectively, both P < 0.05) . After 8-hour treatment with calcofluor white-labeled A. fumigatus spores, confocal laser scanning fluorescence microscopy showed that LC3 and Rubicon mainly surrounded A. fumigatus, suggesting their colocalization with A. fumigatus. Conclusion:A. fumigatus can in vitro increase the basal autophagic flux in murine BMDM.

Microtubular Dysfunction and Male Infertility

Role of LC3 Ⅱ in hyperoxia-induced apoptosis in alveolar type Ⅱ epithelial cells / 中华麻醉学杂志

Objective To investigate the role of microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ) in hyperoxia-induced apoptosis in alveolar type Ⅱ epithelial cells.Methods A549 cells were seeded in 6-well culture plates at a density of 1 × 105 cells/ml (0.5 ml/well) and were divided into 3 groups (n =12 each) using a random number table method:control group (group C),95％ oxygen group (group 95％ O2) and 95％ oxygen plus autophagy inhibitor 3-methyladenine (3-MA) group (group 95％ O2+3-MA).Cells in group C were cultured in incubators containing 95％ air and 5％ CO2 at 37 ℃.Cells were cultured in sealed chambers flushed with 95％ O2 and 5％ CO2 at 37 ℃ in group 95％ O2 and group 95％ O2+3-MA.The cells in each well were pretreated with 3-MA 5 mmol/L before exposure to 95％ O2 in group 95％ O2+3-MA.When the A549 cells were incubated for 24 h,6 wells in each group were selected to detect the expression of LC3 Ⅱ protein and caspase-3 using Western blot,and the left 6 wells in each group were selected to calculate the cell mortality rate by trypan blue dye.Results Compared with group C,the expression of LC3 Ⅱ and caspase-3 was significantly up-regulated,and the cell mortality rate was increased in group 95％ O2 and group 95％ O2+3-MA (P＜0.05).Compared with group 95％ O2,the expression of LC3 Ⅱ was significantly down-regulated,the expression of caspase-3 was up-regulated,and the cell mortality rate was increased in group 95％ O2 and group 95％ O2+3-MA (P＜0.05).Conclusion The up-regulated expression of LC3 Ⅱ can inhibit hyperoxia-induced apoptosis in alveolar type Ⅱ epithelial cells.

Role of LC3Ⅱ in hyperoxia-induced apoptosis in alveolar typeⅡ epithelial cells / 中华麻醉学杂志

Objective@#To investigate the role of microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ) in hyperoxia-induced apoptosis in alveolar typeⅡ epithelial cells.@*Methods@#A549 cells were seeded in 6-well culture plates at a density of 1×105 cells/ml (0.5 ml/well) and were divided into 3 groups (n=12 each) using a random number table method: control group (group C), 95% oxygen group (group 95% O2) and 95% oxygen plus autophagy inhibitor 3-methyladenine (3-MA) group (group 95% O2+ 3-MA). Cells in group C were cultured in incubators containing 95% air and 5% CO2 at 37 ℃.Cells were cultured in sealed chambers flushed with 95% O2 and 5% CO2 at 37 ℃ in group 95% O2 and group 95% O2+ 3-MA.The cells in each well were pretreated with 3-MA 5 mmol/L before exposure to 95% O2 in group 95% O2+ 3-MA.When the A549 cells were incubated for 24 h, 6 wells in each group were selected to detect the expression of LC3Ⅱ protein and caspase-3 using Western blot, and the left 6 wells in each group were selected to calculate the cell mortality rate by trypan blue dye.@*Results@#Compared with group C, the expression of LC3Ⅱ and caspase-3 was significantly up-regulated, and the cell mortality rate was increased in group 95% O2 and group 95% O2+ 3-MA (P<0.05). Compared with group 95% O2, the expression of LC3Ⅱ was significantly down-regulated, the expression of caspase-3 was up-regulated, and the cell mortality rate was increased in group 95% O2 and group 95% O2+ 3-MA (P<0.05).@*Conclusion@#The up-regulated expression of LC3Ⅱ can inhibit hyperoxia-induced apoptosis in alveolar typeⅡ epithelial cells.

Relationship between plasma neurodegenerative protein level and non-motor symptoms in PD patients / 中华老年心脑血管病杂志

Objective To study the relationship between plasma neurodegenerative protein level and non-motor symptoms(NMS)in PD patients.Methods Eighty-four PD patients served as a PD group and 54age-matched persons undergoing physical examination served as a control group. The NMS of PD patients were assessed according to the HAMD scale.The plasma levels of tau,p-tau181,Aβ-42andα-syn were measured by ELISA and analyzed by Spearman correlation analysis and binary logistic regression analysis respectively.Results The FSS score and plasmaα-syn level were significantly higher while the plasma Aβ-42level was significantly lower in PD group than in control group(3.22±1.68 vs 1.89±1.16,P=0.000;320.00±64.91ng/L vs 277.78±52.75ng/L,P=0.000;267.61±77.75ng/L vs 321.80±49.41ng/L,P=0.001).No significant difference was detected in plasma tau and p-tau181levels between the two groups(P＞0.05).The plasmaα-syn level was positively related with the FSS score(r=0.237,P=0.030)and was an influencing factor of FSS(OR=1.019,95％CI:1.006-1.032,P=0.004).Conclusion Plasma neurodegenerative protein level is related with NMS and plasmaα-syn level is a peripheral biomarker for fatigue in PD patients.

Expression and clinical significance of autophagy-related proteins LC3, P62 and Beclin-1 in hypertrophic scar formation / 中国医师杂志

Objective To explore the differences in autophagic expression levels between hypertrophic scar (HS) tissue and normal skin tissue,and further investigate the relationship between hypertrophic scar formation and autophagy protein expression through the rabbit ear hypertrophic scar model.Methods 30 patients with hypertrophic scar were collected.One hypertrophic scar tissue and one normal skin tissue were harvested.The relative expressions of LC3,P62 and Beclin-1 in each tissue specimen were detected by immunohistochemistry and Western blot.Western blot was used to detect the autophagic-associated protein LC3 (MAPLC3),P62 and Beclin-1 in the hypertrophic scar tissue of rabbit ear and the corresponding normal tissue of rabbit ears at 4 weeks,8 weeks,12 weeks,and 24 weeks,and further explore their clinical significance.Results In vivo,the expression of hypertrophic scar tissue protein LC3 and Beclin-1 was significantly stronger than that in normal skin tissue (P ＜ 0.05).The expression of P62 was significantly weaker than that in normal skin tissue (P ＜ 0.05).In animal experiments,during the process of HS formation,the protein expression of LC3 gradually increased,while the protein expression of P62 gradually decreased;the protein expression of Beclin-1 was higher than that of normal rabbit ears tissue,with statistically significant differences (P ＜ 0.05).Conclusions The expression of LC3 and Beclin-1 in human hypertrophic scar tissues is higher than that in normal tissues.While the expression of P62 is lower than that in normal tissues.That is,the expression of autophagy in human hypertrophic scar tissue showed an upward trend in a certain period of time,and was significantly higher than that in normal tissue.

In vitro effect of low-concentration hydrogen peroxide on autophagy in human melanocytes and screening for autophagy-related lncRNAs / 中华皮肤科杂志

Objective To evaluate the effect of the treatment with low-concentration hydrogen peroxide (H2O2) on the adhesive function of and autophagy in human melanocytes,and to screen long noncoding RNAs (lncRNAs) related to autophagy.Methods Melanocytes were isolated from foreskins of healthy males after circumcision,and subjected to cultivation.Melanocytes at exponential growth phase were divided into 3 groups:control group receiving no treatment,H2O2 group treated with 400 μ mol/L H2O2,and H2O2 + NAC group pretreated with 4 mmol/L NAC for 2 hours followed by the treatment with 400 μmol/L H2O2.After 5-day treatment,immunofluorescence study was performed to determine the expression of Ecadherin,microtubule-associated protein 1 light chain 3 (LC3)and p62,and Western blot analysis to determine the expression of autophagy-related protein LC3 and p62.Cell structures and autophagosomes were observed by transmission electron microscopy,and autophagy-related lncRNAs were screened using gene chip technology.Statistical analysis was done with Graphpad Prism 6 software using one-way analysis of variance for comparison among groups,and Tukey's test for multiple comparisons.Results Under the confocal microscopy,the H2O2 group showed significantly decreased fluorescence intensity of E-cadherin and LC3 in the melanocytes and decreased number of autophagosomes in melanocytes,but significantly increased fluorescence intensity of p62 compared with the control group and H2O2 + NAC group.Western blot analysis showed that the LC3-Ⅱ/LC3-Ⅰ ratio in the melanocytes was significantly lower in the H2O2 group (0.604 ± 0.012) than in the control group (1.200 ± 0.081,q =7.718,P ＜ 0.01) and H2O2 + NAC group (1.017 ± 0.062,q =5.076,P ＜ 0.05),while the p62/β-actin ratio in the melanocytes was significantly higher in the H2O2 group (0.881 ± 0.079) than in the control group (0.456 ± 0.121,q =4.847,P ＜ 0.05) and H2O2 + NAC group (0.492 ± 0.049,q =4.439,P ＜ 0.05).There were no significant differences in the LC3-Ⅱ/LC3-Ⅰ ratio or p62/β-actin ratio between the H2O2 + NAC group and control group (P ＞ 0.05).Gene chip technology showed that 18 autophagy-related lncRNAs were associated with premature senescence of melanocytes and differentially expressed in the H2O2 group compared with the control group,and the autophagy-related lncRNA NONHSAT190308.1 (＞ 10-fold increase) was screened out.Conclusion Lowconcentration H2O2 can decrease the expression of E-cadherin and the level of autophagy in melanocytes,and can up-regulate the expression of autophagy-associated lncRNA NONHSAT190308.1.

The role of UCH-L3 and LC3Ⅱin the diagnosis of acute myeloid leukemia / 中华检验医学杂志

Objective Todetect the mechanism of acute myeloid leukemia ( AML ) and the relationship between autophagy andubiquitin c-terminal hydrolases L3( UCH-L3) for providing new targets and guiding clinical management in AML .Methods 84 cases of AML patients and 25 controls were chosen from the First Affiliated Hospital of Dalian Medical University from 2014 to 2015,including 47 males and 37 females.The AML patients were divided into 3 groups :initialdiagnosis group [40 cases including 24 males and 16 females,with an median age of 54 (23 -85)];complete remission group [30 cases including 14 males and 16 females,with an median age of 45 (22-74)]and refractory group[14 cases including 9 males and 5 females,with an median age of 50 (18-80)].Among those patients, there were 3 cases of M1, 42 cases of M2, 18 cases of M3, 3 cases of M4 and 18 cases of M5 subtypes.The expression of UCH-L3 and LC3II were detected by Real Time PCR and Western blot after the HL-60 cells were treated by TCID.The expression of UCH-L3 and LC3IIin the PBMC of AML patients and controls were detected with Real Time PCR.The expression levels of UCH-L3 and LC3II in different types of AML were analyzed .The relationship between the expression of LC 3Ⅱand clinical features , clinical stages , laboratory results and therapeutic effects of patients were investigated .It further detected the expression of UCH-L3 and LC3IImRNA in post-induction status.t test were used for measurement data , χ2 test were performed for rate comparison; rank correlation test were performed for correlation analysis .Non-parametric test were performed for non-normal distribution data.Results Both the UCH-L3 and LC3 II were down-regulated after TCID treatment in HL-60 cellsat gene and protein levels (t=-29.435, t=-8.105,P<0.05).The expression of UCH-L3 in initial diagnosis group was lower than control group ( Z=-3.87,P<0.05),butit was higher in remission group than initial diagnosis group and refractory group ( Z=-6.70 , Z=-4.09, P<0.05 ) .The expression level of LC3Ⅱin diagnosis group was significantly higher than control group , remission group and refractory group(Z=-6.96,Z=-5.32, Z=-3.52,P<0.05).Cytogenetic abnormalities in patients with high expression of LC3II were more common(χ2 =6.510,P<0.05).The relative expression of UCH-L3 and LC3ⅡmRNA in the same patient before and after treatment were significantly different ( P<0.05 ) . During the remission, the expression of UCH-L3 was up-regulated compared with that before primary treatment, while the expression of LC3II was down-regulated.There was no statistically significant in the relative expression of UCH-L3 and LC3Ⅱamong the FAB type.Conclusion The UCH-L3 and autophagy are associated with pathogenesis of AML .

Role of Microtubule-Associated Protein in Autism Spectrum Disorder / 神经科学通报·英文版

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by deficits in social interaction and communication, along with repetitive and restrictive patterns of behaviors or interests. Normal brain development is crucial to behavior and cognition in adulthood. Abnormal brain development, such as synaptic and myelin dysfunction, is involved in the pathogenesis of ASD. Microtubules and microtubule-associated proteins (MAPs) are important in regulating the processes of brain development, including neuron production and synaptic formation, as well as myelination. Increasing evidence suggests that the level of MAPs are changed in autistic patients and mouse models of ASD. Here, we discuss the roles of MAPs.

Expression and significance of autophagy-related genes in acute pancreatitis / 中华胰腺病杂志

Objective To examine the expression and analyze the significance of autophagy-related gene microtubule-associated protein 1 light chain 3 (MAP1-LC3,LC3),p62 and lysosorne-associated membrane protein 2 (LAMP-2) in pancreatic tissues of mice with acute pancreatitis (AP).Methods Twenty mice were randomized into AP group and control group,and the number of mice was equal between two groups.AP group was intra-peritoneally injected by 20％ L-arginine solution (two injections of 4 g/kg body weight,every 1 h) in the dosage of 4 g/l kg twice every 1 hour to establish AP model,while control group was administered with equal volume of normal saline by intra-peritoneal injection.All the mice were euthanized at 24 hour after the last injection.Pancreatic histopathological changes were measured.In addition,the protein expressions of LC3,p62 and LAMP-2 were detected by Western blot.Results No obvious pathological changes were observed in control group.Pancreatic acinar edema,structure destruction,missing,the obvious widening of interlobular septum,small interlobular septum and acinar septum,and the necrosis of acinar cells at different degrees were observed in AP group.The pathological score for tissue edema,hemorrhage,necrosis and inflammation in AP group was 3.13 ± 0.50,2.83 ± 0.32,3.25 ± 0.46 and 3.16 ± 0.47,respectively,which was all 0 in control group.The differences were statistically significant between AP group and control group (P ＜ 0.01).In AP group,the ratio of LC3-Ⅱ/LC3-Ⅰ,p62 and LAMP-2 protein in pancreatic tissue were 1.16 ± 0.08,0.94 ± 0.04 and 0.35 ± 0.04,respectively,which were 0.24 ± 0.02,0.34 ± 0.03 and 0.95 ± 0.03 in control group.The ratio of LC3-Ⅱ/LC3-Ⅰ and p62 protein in pancreatic tissue in AP group were much higher than those in control group,while LAMP-2 in AP group was lower than that in control group,and there was statistically significant difference between two groups (all P ＜0.01).Conclusions Intraperitoneal injection of L-arginine could induce acute pancreatitis,and autophagy is impaired,which was associated with decreased LAMP-2 protein expression.

Effects of PI3K / Akt signaling pathway on the autophagy of rat alveolar macrophages exposed to dust / 中国职业医学

OBJECTIVE: To determine the regulating role of phosphatidylinositol 3-kinase( PI3K) / protein kinase B( Akt)signaling pathway in the autophagy activity of rat NR8383 cells exposed to silicon dioxide( SiO_2). LY294002 was used to block PI3 K pathway. METHODS: i) The normal NR8383 cells were used and divided into blank group and silica exposure group( final concentrations of SiO_2 suspension were 0 and 50 mg / L respectively). They were cultured for 3,6,12,20 and24 hours. The enzyme linked immunosorbent assay( ELISA) was used to assess the amount of tumor necrosis factor-α( TNF-α) and transforming growth factor-β1( TGF-β1) in supernatants of cultured cells,and then the optimal time of cells exposed to dust was determined. ii) NR8383 cells were divided into control group( treated with a same volume of F-12 K medium without serum),silica group( treated with SiO_2 suspension,final concentration 50 mg / L) and intervention group( treated with SiO_2 suspension and PI3 K inhibitor LY294002,final concentration 50 mg / L and 20 μmol / L,respectively).Cells were harvested following incubation. ELISA was used to detect the levels of TNF-α and TGF-β1 at the time point of20 hours after incubation. To reveal the autophagy status of cells,Western blotting was used to detect Akt and microtubuleassociated proteins 1 light chain 3( LC3) protein at time point of 20 hours; laser scanning confocal microscope( LSCM)was used to observe the immunofluorescence expression of autophagy at time points of 3,6,12 and 20 hours. The cells were also treated with the lysosomal inhibitor chloroquine diphosphate( CDP) at the same time of SiO_2 treatment. RESULTS: i) The time point of 20 hours was confirmed to be the best dust exposure time for in vitro cell model of NR8383 cells.ii) The levels of TNF-α and TGF-β1 of supernatant in the silica group were higher than those of the control group( P <0. 05). The levels of TNF-α and TGF-β1 of supernatant in the intervention group were higher than those of the control group and silica group( P < 0. 05). The Akt protein expression of the intervention group was lower than those in the control group and the silica group,respectively. The LC3 Ⅱ / Ⅰ protein level of the silica group was higher than those of the control group and intervention group( P < 0. 05),but no statistical significance was found between the control group and intervention group( P > 0. 05). LSCM results indicated that autophagy expression at time points of 3 and 6 hours were stronger than those of 12 and 20 hours in control group; autophagy expression at time point of 12 hours was stronger than those of 3 and 6 hours in the silica group,while the autophagy expression at time point of 20 hours was slightly weaker than that of 12 hours,but still stronger than those of 3 and 6 hours. Compared with the same time point in control group,autophagy expression at 3 and 6 hours were weaker in the silica group,while the expressions increased obviously at time points of 12 and 20 hours. Autophagy expression at all time points decreased in the intervention group compared with silica group,especially at the time point of 20 hours. The autophagy expression in each group increased in varying degrees after added with CDP blocking. CONCLUSION: Silica dust exposure can induce autophagy in rat NR8383 cells. PI3 K inhibitor LY294002 can reduce the autophagy expression indicating that the PI3 K / Akt signaling pathway might participate in the autophagy process of silica dust inducing autophagy in alveolar macrophages.

The change of phosphorylated MAP-2 in hippocampus/amygdala and the influence of ginsenoside Rb1 on it in depressive rat model / 天津医药

Objective To observe effects of ginsenoside Rb1 on phosphorylation of microtubule-associated protein 2 (pMAP-2) in hippocampus and amygdala of depressive model rats. Methods Thirty male Wistar rats were randomly divided into control group, model group and treatment group. The depression rat model was produced by giving chronic unpredicted mild stress (CUMS). Treatment group was given daily intragastric administration of ginsenoside RB 1 (1 g/mL crude drug, 1 mL/100 g body weight) for 22 days during modeling. Western blot assay was used to detect expressions of MAP-2 and pMAP-2 protein, and real-time PCR was used to detect expressions of pMAP-2 mRNA respectively. Results The expressions of pMAP-2 protein and mRNA in hippocampus and amygdala were significantly lower in model group than those of control group (P < 0.05). The expressions of pMAP-2 protein and mRNA were significantly higher in treatment group than those of model group (P<0.05). Conclusion Ginsenoside Rb1 can play anti-depression role by inhibiting the phosphorylation of MAP-2 in rats.

Improvement effect of BQ-123 on nerve function damage after subarachnoid hemorrhage in rats / 吉林大学学报(医学版)

Objective:To study the effect of endothelin receptor antagonist BQ-123 on the nerve function damage after subarachnoid hemorrhage (SAH)in the rats,and to explore the mechanisms.Methods:Total 120 male SD rats were divided into sham group,SAH group,low dose of BQ-123 group (50 μg· kg-1 )and high dose of BQ-123 group (75 μg·kg-1 ).The SAH rat models were established by injecting the autologous blood into cisterna magna twice.The morphological changes of hippocampus nerve cells of rat brain tissue were detected with HE staining, and the expressions of mTOR, Beclin-1 and LC3-Ⅱ in the hippocampus of rats were detected with immunohistochemistry and RT-PCR;the shuttle-box experiment was used to evaluate the abilities of learning and memory,and the holding power evaluation was used to evaluate the forelimb pulling force of the rats in various groups at each time point.Results:Compared with sham group,the morphological damages of neurons of the rats in SAH group were increased,the survival rate of neurons of the rats in SAH group was decreased (P <0.05),the expression levels of mTOR mRNA,Beclin-1 mRNA and LC3-Ⅱ mRNA in hippocampus tissue of the rats were increased (P < 0.05),and the abtilities of learning and memory and the values of holding power were decreased (P <0.05).Compared with SAH group,the morphological damages of neurons of the rats in BQ-123 groups were decreased,the survival rates of neurons of the rats in BQ-123 groups were increased (P < 0.05),the expression levels of mTOR mRNA of rats were decreased (P <0.05),the expression levels of Beclin-1 mRNA and LC3-ⅡmRNA in hippocampus tissue were increased (P <0.05),and the abilities of learning and memory and the values of holding power were increased (P < 0.05 ). The changes were more significant in high dose of BQ-123 group compared with low dose of BQ-123 group (P <0.05).Conclusion:BQ-123 can improve the nerve function damage after SAH in the rats,its mechanism may be related to regulating the mTOR/autophagy signaling pathway.

Expression of doublecortin-like kinase 1 in colorectal cancers and colorectal adenomas and its clinical significance / 中国医师杂志

Objective To investigate the expression of doublecortin-like kinase 1 (DCLK1) in colorectal cancer and colorectal adenomas,and it's role in the development and progression of colorectal carcinoma,and explore the relationship of the DCLK1 expression and prognosis in colorectal cancer patients with diabetes medical history.Methods Expression of DCLK1 in 70 cases of colorectal cancers,70 cases of para-carcinoma tissues,and 35 cases of tubulovillous adenomas were detected by immunohistochemical method.Results (1) The DCLK1 high expression rate in colorectal cancers was 41.4％ (29/70),and 45.7％ (16/35) in tubular villous adenomas,both significantly higher than the para-carcinoma tissues (4.3％;3/70) (P ＜0.01).(2) DCLK1 high positive expression had statistical significance with pathological grading (P ＜ 0.01),infiltration depth (P ＜ 0.05),lymph node metastasis (P＜ 0.05),vascular invasion (P ＜0.05),and distant metastasis (P ＜0.01).(3) There was no obvious correlation between the Diabetes medical history,expression of DCLK1,and prognosis of patients.Conclusions DCLK1 has high positive expression in colorectal carcinomas,and may have certain significance for the early clinical diagnosis and prognostic judgment of colorectal cancers.The expression of DCLK1 and prognosis of colorectal cancers has no obvious correlation with diabetes medical history.

Expression and role of microtubule-associated protein in breast cancer / 国际肿瘤学杂志

In recent years,many new microtubule-associated proteins are found or studied extensively in the treatment of breast cancer.End-binding protein 1 (EB1)has the promotional effects on cell proliferation and enhances the carcinogenesis of breast cancer,and also can affect the chemosensitivity of the treatment. MAP7 domain-containing protein 3 (Mdp3)expresses in breast epithelial cells and plays an important role on the growth and metastasis.Nucleolar and spindle-associated protein (NuSAP)gene is up-regulated in breast cancer and is considered as a biomarker for breast cancer.Tau expression level has a strong correlation with estrogen receptor (ER),progesterone receptor (PR)and human epidermal growth factor receptor-2 (HER-2), and it remains controversial in the results of the effect of paclitaxel chemotherapy and hormone treatment.

α-Galactosidase A gene mutation induced autophagic dysfunction and its mechanism / 中华神经科杂志

Objective To investigate the effect of α-galactosidase A (GLA) gene mutation on cell autophagy and to elucidate its mechanism preliminarily.Methods Two families were diagnosed by ultrastructural pathological examination,GLA gene activity test and GLA gene mutation screening.Mutant type recombinant expression plasmid of two pedigrees (pcDNA3.1-GFP-ex1 (EX1 group),pcDNA3.1-GFP-ex3 (EX3 group)) and wild type recombinant expression plasmid of GLA (pcDNA3.1-GFP-GLA,GLA group) were constructed.Hela cell line (control group) was transiently transfected with recombinant expression plasmid according to lipofectin transfection.The relative gene expression of Beclin-1 was measured with real-time PCR,and protein expression level of LC3-Ⅱ/LC3-Ⅰ,Beclin-1 and P62/SQSTM1 was examined by Western blotting.Results The LC3 protein values of groups EX1,EX3,GLA and control were 1.495 ± 0.064,1.490 ± 0.020,1.285 ± 0.021,1.260 ± 0.042,respectively;P62/ SQSTM1 values were 0.555 ± 0.086,0.480 ± 0.084,0.785 ± 0.439,0.980 ± 0.278,respectively;Beclin-1 mRNA 2-△Ct values were 0.011 ±0.003,0.008 ±0.002,0.005 ±0.001,0.003 ±0.001,respectively;Beclin-1 protein values were 1.178 ±0.098,1.209 ±0.092,0.931 ±0.100,0.796 ±0.184,respectively.Compared with the wide type group,the level of LC3-Ⅱ/LC3-Ⅰ protein was significantly higher in the mutant type groups(t =5.118,4.984;P =0.007,0.008),though no statistically significant difference was found in the expression levels of P62/SQSTM1 (t =1.052,1.400;P =0.323,0.199).Besides,the expression levels of Beclin-1 mRNA (t =3.800,2.445;P =0.005,0.040) and protein (t =2.424,2.729;P =0.042,0.026) were significantly higher in the mutant type groups.Conclusions GLA gene mutation can induce cell autophagic dysfunction,and signaling pathway of autophagic activation may be Beclin-1 dependent.

Expression and clinical significance of CIP2A, PI3K, and survivin in pancreatic carcinomas / 中国医师杂志

Objective To investigate the expressions of cancerous inhibitor of protein phosphatase2A (CIP2A),phosphatidylinositol 3-kinase(PI3K),and survivin in pancreatic carcinomas and their clinical significance.Methods The expressions of CIP2A,PI3K,and survivin proteins were tested by immunohistochemistry in 64 cases of pancreatic carcinomas and adjacent paracancerous tissues.Results The positive rate of CIP2A in pancreatic carcinomas was significantly higher than adjacent paracancerous tissues (70.3％ vs 5.6％,P ＜0.05).Significant difference was observed in the expression rate of PI3K between the patients with pancreatic carcinomas and paracancerous tissues (73.4％ vs 8.3％,P ＜0.05).Significant difference was also observed in the expression rate of survivin between the patients with pancreatic carcinomas and paracancerous tissues (75.0％ vs 2.8％,P ＜0.05).CIP2A,PI3K,and survivin were significantly differentially expressed in pancreatic carcinoma among different tumor differentiation,tumor node metastasis (TNM) stage,and neural invasion and lymph node metastasis (all P ＜ 0.05).Spearman correlation analysis showed significantly positive correlation between the expressions of CIP2A and the others (PI3K and survivin) (both P ＜0.05),and between the expressions of PI3K and surviving (P ＜0.05).Conclusions CIP2A was involved in the development of pancreatic carcinomas and might activate the PI3K/Akt/survivin pathway.Our data identified CIP2A as a critical oncoprotein involved in cell proliferation,invasion,and metastasis.It could serve as a therapeutic target for pancreatic carcinomas.

Expressions and their Significance of Ki-67, p53, and survivin in esophageal cancer and precancerosis / 中国医师杂志

Objective To investigate the expressions and clinical significance of Ki-67,p53,and survivin in esophageal cancer and precancerosis.Methods The expressions of Ki-67,p53,and survivin proteins were detected by immunohistochemical staining in 40 normal esophageal mucosa,136 precancerosis (42 mild atypical hyperplasia,43 moderate atypical hyperplasia,and 51 severe atypical hyperplasia),and 68 esophageal cancer tissues.The correlation of three proteins expressed in esophageal carcinoma tissues was analyzed.Results The positive expression rate of Ki-67 was 0 (0/40)for normal epithelium,35.7％ (15/42) for mild dysplasia,51.2％ (22/43) for moderate dysplasia,74.5％ (38/51) for severe dysplasia,92.6％ (63/68) for squamous carcinoma,respectively.The positive expression rate of p53 protein was 0 (0/40) for normal epithelium,28.6％ (12/42) for mild dysplasia,46.5％ (20/43) for moderate dysplasia,52.9％ (27/51) for severe dysplasia,67.6％ (46/68) for squamous carcinoma,respectively.The positive expression rate of survivin protein was 0 (0/40) for normal epithelium,38.1％ (16/42) for mild dysplasia,55.8％ (24/43) for moderate dysplasia,64.7％ (33/51) for severe dysplasia,89.7％ (61/68) for squamous carcinoma,respectively.Rank correlation analysis showed that abnormal expressions of Ki-67,p53,and survivin were correlated significantly with the pathological grading of the lesions (r =0.637,0.454,0.590,P ＜0.01).The expressions of Ki-67,p53,and survivin were positively correlated in esophageal carcinoma (r =0.407,0.646,P ＜ 0.01).Conclusions The abnormal expressions of Ki67,p53,and survivin were associated with the processes of the esophageal canceration,and the joint detection with three parameters has important clinical value.

Expressions and clinical significance of survivin and lymphatic vessel endothelial hyaluronan recep-tor 1 in gastric carcinoma / 中国医师杂志

Objective To investigate the expressions and clinical significance of survivin, and lymphatic vessel endothelial hyaluronan receptor 1 ( LYVE-1) in gastric cancers.Methods The expres-sions of survivin and LYVE-1 were detected with streptavidin avidin-peroxidase ( SP) immunohistochemical method in 70 cases of gastric cancers and 50 cases of normal gastric tissues.The relationship between the expressions of survivin and LYVE-1 with the clinicopathological parameters was analyzed.Results The ex-pression rate of survivin was 65.71%(46/70) in gastric cancers, and 6.00% (3/50) in normal gastric tissues, with statistically significant difference between two groups (χ2 =43.048, P 0.05).The expression rate of LYVE-1 was 32.86%(23/70) in gastric cancers, and 4.00%(2/50) in gastric ulcers, with significant difference between two groups (χ2 =14.726, P 0.05 ) .There was a positive correlation between expressions of survivin and LYVE-1 in gastric cancers ( r =0.271, P <0.05).Conclusions Survivin and LYVE-1 were abnormally expressed in gastric cancers with a colose relationships between two parameters.They might play important roles in tumorigenesis and progression of a gastric cancer.