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Objective To explore the influence of apoptosis-associated speck-like protein containing a caspase recruitment do-main(ASC)and Caspase-1 on the pathogenesis of primary biliary cirrhosis(PBC).Methods The real-time PCR,Western blot,py-rophosphate sequencing and ELISA were adopted to respectively detect the relative expressions of mRNA and protein of peripheral blood mononuclear cells(PBMS)Caspase-1 and ASC as well as the methylation status of ASC promoter region and plasma Caspase-1 and IL-18 expression levels in 30 cases of PBC and healthy controls.Results The mRNA and protein expressions of PBMC Caspase-1 and ASC in the PBC group were significantly higher than those in the control group(P<0.05).The methylation rate of ASC promoter Island1(ISI)was significantly lower than that of the control group(P<0.05),which of Island 2(IS2)was smaller than the background value and had no methylation occurrence.The levels of plasma Caspase-1 and IL-18 in the PBC group were sig-nificantly higher than those in the control group(P<0.05).The ASC mRNA in the PBC group was significantly correlated with the Caspase-1 mRNA expression(P<0.05);the methylation rates at loci 1,2,4,5 of ASC promoter region CpG island were nega-tively correlated with ASC mRNA expression(P< 0.05),and which at loci 3,6 had no correlation with their expressions(P>0.05);plasma Caspase-1 and IL-18 levels showed the obviously positive correlation.Conclusion ASC and Caspase-1 are involved in the immune inflammatory response in PBC.
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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 .
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Objective To investigate the effect of mild hypothermia on the expression of small ubiquitin-like modifier-specific proteases 3 (SENP3) in the brain tissues during cerebral ischemia-reperfusion (I/R) in mice.Methods Ninety-six male C57/BL6 mice,aged 10-12 weeks,weighing 22-30 g,were randomly divided into 3 groups (n =32 each) using a random number table:sham operation group (group S),group I/R,and mild hypothermia group (group H).Cerebral I/R was produced by occlusion of bilateral common carotid arteries for 15 min followed by reperfusion.The surface cooling was started immediately after reperfusion,and the rectal temperature was maintained at 32-34 ℃ for 3 h.At 6,12,24 and 72 h of reperfusion,8 mice were selected from each group and sacrificed.The hippocampi were removed for examination of the pathological changes (with light microscope) and for determination of cell apoptosis (using TUNEL) and expression of SENP3 (by Western blot).The apoptosis rate was calculated.Results Compared with group S,the apoptosis rate in hippocampal CA1 region was significantly increased,and the expression of SENP3 was significantly up-regulated at each time point of reperfusion in group I/R (P＜0.05).Compared with group I/R,the apoptosis rate in hippocampal CA1 region was significantly decreased,and the expression of SENP3 was significantly down-regulated at each time point of reperfusion (P＜0.05),and the pathological changes were significantly reduced in group H.Conclusion The mechanism by which mild hypothermia reduces cerebral I/R injury is associated with inhibition of SENP3 expression in the brain tissues of mice.
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Objective To study the effects of Liensinine on apoptosis of 5637 cells, and its mechanism thereof. Meth?ods CCK-8 method and the colony formation test were used to detect cell viabilities, and then inhibition rates were calcu?lated. Flow cytometry was used to detect the effects of Liensinine on apoptosis of 5637 cells. Western blot assay was used to detect Caspase-7 protein expression. Results CCK-8 assay and colony formation test indicated that Liensinine inhibited the cell proliferation significantly. Results of flow cytometry indicated that Liensinine induced early apoptosis of 5637 cells. Western blot assay showed that Liensinine improved the expression of Caspase-7 and enhanced the activation of Caspase-7 in 5637 cells. Conclusion Liensinine could inhibit the proliferation of 5637 cells, induce early apoptosis, which may be re?lated with the enhanced expression of Caspase-7 and its activation.
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Objective To investigate the occurrence law of autophagy in trophoblast cells from preeclampsia and its underlying mechanisms.Methods Twenty cases of placenta tissues were collected from women suffered from preeclampsia and normal pregnant women respectively.Autophagosome of trophoblast cells were observed by transmission electron microscope.The expressions of LC3-Ⅱ/Ⅰ and Atg4B in placenta tissues were detected by western blot and real-time PCR.Results Compared with the control group,typical autophagosomes of trophoblast cells were observed by transmission electron microscope.The ratio of LC3-Ⅱ / Ⅰ in placenta of PE patients was increased (1.43 ± 0.23) compared with control group (0.59 ±0.12),and the expression of Atg4B was up-regulated in both mRNA [(1.73 ±0.16) folds] and protein levels (0.71 ± 0.13) compared with control group (P ＜ 0.05).Conclusions Autophagy was significantly up-regulated in trophoblast cells from patients suffered from preeclampsia.Thus,all the data suggest that autophagy might be involved in the generation of preeclampsia.
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Objective To investigate the expression of low molecular mass polypeptide-2 (LMP2)and protein phosphatase 1A (PPM1A) in gestational trophoblastic disease and elucidate their predictive value in malignant transformation of hydatidiform mole. Methods The expressions of LMP2 and PPM1A protein in 196 complete hydatidiform moles (in which 28 cases with malignant transformation) , 7 invasive moles, 5 choriocarcinomas and 20 normal chorionic villus were detected with the method of En Vision immunohistochemistry. Their clinicopathologic data were retrospectively analyzed. Results LMP2 and PPM1A protein expressed in cytotrophocytes, syncytiotrophoblast and extravillous trophoblast. The level of LMP2 expression in deteriorative hydatidiform mole was significantly higher than that in non-deteriorative hydatidiform mole or normal chorionic villus (6. 79 ±2. 38, 5.26 ±2.63 and 3. 10 ±1.65, all P 0. 05). Conclusions High expression of LMP2 and low expression of PPM1A might play an important role in the motility and invasiveness of trophohlast cells and malignant transformation of hydatidiform mole. Testing the expression of LMP2 and PPM1A in hydatidiform mole tissues of initial uterine evacuation might be have some reference significance in judging outcomes of hydatidiform mole.
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OBJECTIVE To investigate the inhibiting effect of Pingyangmycin on cultured vein endothelial cells in vitro and illustrate the mechanism of Pingyangmycin on treatment cavernous hemangioma of head and neck. METHODS By MTT assay,the inhibiting rates of cultured Ecv-304 cells which were managed with Pingyangmycin for 24h and 48h were compared respectively. The changes in the cell cycle,apoptosis,and Caspase 3 protein expression of the cells managed with Pingyangcin for 24h were examined by flow cytometry(FCM). RESULTS The results showed that the inhibiting rate of Ecv-304 was dependent on the concentration of Pingyangmycin. However,in the same concentration,the inhibiting effect for 24h was stronger than that for 48h. The percentage of G1 phase cells increased while the S phase decreased,but the percentage of G2 phase cells remained unchanged after managed with Pingyangcin for 24h. An apoptosis peak appeared before G1 phase. The apoptosis rate was also concentration-dependant. The expression of caspase 3 increased in every group of different concentration. CONCLUSION Pingyangmycin can possibly arrest the growth of Ecv-304 in the G1 phase through the mechanism of inducing apoptosis. Caspase 3 may play an important role in the induced apoptosis.