Objective To compare the influence of aging on apoptosis and the expression of apoptosis-related protein in adult and aged male Wistar rat hearts after acute coronary artery occlusion. Methods The acute myocardial infarction rat model was established by left anterior descending(LAD)occlusion.A total of 115 adult and aged rats,aged 6-24 months,were included in the study.The rats were divided into 4 groups:aged model group,aged control group,adult model group and adult control group.Animals were killed 1,3,5 hours,1 and 7 days after coronary occlusion.Hemodynamic parameters[heart rate(HR),left ventricular systolic pressure(LUSP),left ventricular end-diastolic pressure(LVEDP),±dp/dtmax)]were obtained from each group at every time points.The apoptosis and necrosis of myocardium were detected with TUNEL way and TTC stain.The expressions of Bcl-2 and Bax were analyzed with immumohistochemical stain. Results DNA fragmentation occured 1 hour after coronary occlusion and apparently peaked earlier in the aged than in the adult rat hearts.At 3 hour,the apoptotic index of aged model group was obviousily higher than that in adult model group[(51.90±23.15)%us.(18.67±17.15)%,P<0.01].The basal levels of Bcl-2 and Bax were higher in the aged than in the adult rat hearts.The expression of Bcl-2 in aged model group and adult model group were 2.7±0.9 and 1.8±0.8,P<0.05.The expression of Bax in aged model group and adult model group were 6.2±2.9 and 4.2±1.5,P<0.05. Conclusions The ability of aged rats to resist ischemia is poor,aging may alter the expressions of Bcl-2 and/or Bax,increase cardiomyocyte apoptosis,thereby,enhance the myocardial dysfunction during acute myocardial infarction.

Objective To observe the translations of chemokine CXCL12 and its receptor CXCR4 in endometrial carcinoma tissues,and to evaluate their significance in clinical pathologic features of endometrial carcinoma.Methods From Jan.2012 to Dec.2014,52 tissue specimens of patients with endometrial carcinoma were admitted to the hospital,aged 55.6 ± 19.2 years,and 26 cases of normal endometrium as control,aged 52.3±16.5 years old.Expressions of CXCL12 and CXCR4 in the tissue specimens were detected by immunohistochemistry,and to analyze the relationship between the expressions and clini-cal pathological features,and FIGO stage,cell differentiation,lymph node metastasis and pathological type in endometrial carcinoma.Results In endometrial carcinoma,the positivity rates of CXCR4 and CXCL12 were 76.92% and 69.23% re-spectively.The positive expression rates of which were 34.62% and 30.77% in normal endometrium tissues,statistically significant (χ2 =11.826,P <0.01).The expression of CXCR4 was positively correlated with the differentiation of endome-trial carcinoma cells (P <0.05).In endometrial carcinoma,with the exception of CXCR4 expression and cell differentiation was positively related (r=0.386,P <0.05),CXCL12 and CXCR4 expression and clinical stage,pathological type and lymph node metastasis no correlation was found (P >0.05).Conclusion The results showed that the expression of CXCL12 and CXCR4 in endometrial carcinoma tissues was significantly increased,which may play an important role in the development of endometrial cancer.

Humans ; Work

Private Sector ; Public Sector ; Social Responsibility

OBJECTIVE:To study the stability and to predicate the expiration date of betamethasone cream.METHODS:The HPLC method was used to determination the content of betamethasone,the stability test of cream was done by initial mean velocity method and strong light exposure experiment,respectively.RESULTS:The linear concentration range for betametha?sone cream was0.25～8?g/ml(r=0.9991),the average recovery rate was98.56%,RSD=0.78%;The content was highly correlated to temperature,whereas on which the light exposure did little effect.CONCLUSIONS:Betamethasone cream should be stored under room temperature and in a cool place,it is predicted that the expiration date of betamethasone cream is1year.

Objective To investigate the spatial learning and memory ability,the changes of indicators of oxidative stress,and their relationship in transgenic APP/PS1 mouse model of Alzheimer's disease(APP/PS1 mice). Methods The spatial learning and memory ability were assessed by Morris water maze test,and the activity or content of SOD, GSH-PX, MDA, and protein carbonyl in brain tissues were measured by ELISA in the APP/PS1 and wild type (WT) mice. Furthermore, the relationship between the learning and memory performances and the indicators of oxidative stress was examined. Results No significant difference in the spatial learning was observed between the APP/PS1 and WT mice (P ＜0. 05). The spatial memory which was measured as the percentage of time traveling in the targeted quadrant to the total traveling time was significantlydeclined in the APP/PS1 mice(29. 02 ± 4. 27) % as compared with the WT mice(47. 39 ± 6. 01) %(t =0. 000 ,P ＜0. 05). The percentage of length of traveling in the targeted quadrant to the total length traveled was significantly lower in the APP/PS1 mice(28. 85 ±3.77)% compared with the WT mice(46. 70 ±5.60)% (t =0. 000,P ＜0. 05). These findings indicated that the spatial learning and memory ability of APP/PS1 mice was significantly decreased compared to WT mice. There was no significant difference in activity or content of SOD,GSH-PX,and MDA in brain tissues between the APP/PS1 and WT mice (P ＜ 0. 05), while the content of protein carbonyl was significantly elevated in the APP/PS1 mice (2. 67 ±0. 19) than in the WT mice (2. 38 ±0. 15)(t = 0. 008, P ＜ 0. 05). Correlation analysis revealed that the elevated protein carbonyl was negatively correlated with the percentage of length traveled in the targeted quadrant(r = - 0. 639, P ＜ 0. 05) and the percentage of time traveled in the targeted quadrant(r = - 0. 636 ,P ＜ 0. 05). Conclusion The spatial memory impairment was negatively correlated with the elevated protein carbonyl in the APP/PS1 mice, suggesting that protein carbonylation caused by oxidative stress might play an important role in the development of memory impairment in the early stage of Alzheimer's disease.

Objective To investigate the correlation between liver and spleen stiffness measured by acoustic radiation force impulse (ARFI) and hepatic venous pressure gradient (HVPG),and to evaluate its efficiency in the diagnosis of portal hypertension.Methods From April 2014 to March 2016,20 cases underwent HVPG measurement because of liver cirrhosis were enrolled.Before HVPG measurement,liver and spleen stiffness were assessed with ARFI.The correlation between HVPG and age,alanine aminotransferase (ALT),aspartate aminotransferase (AST),total hilirubin,serum albumin,platelet count,prothrombin time,aspartate aminotransferase to platelet ratio index (APRI) score,Child-Pugh score,model for end-stage liver disease (MELD) score,liver stiffness and spleen stiffness were analyzed.Pearson correlation and Spearman rank correlation were performed for statistical analysis.Results HVPG,liver and spleen stiffness were successfully measured in all 20 patients.The mean liver stiffness was (1.78±0.29) m/s,the mean spleen stiffness was (3.37±0.44) m/s and HVPG was (16.10±5.14) mmHg (1 mmHg=0.133 kPa).Age,ALT,AST,total bilirubin,serum albumin,platelet count,prothrombin time,APRI score,Child-Pugh score and MELD score were all not correlated with HVPG (all P＞0.05).But HVPG was positively correlated with liver and spleen stiffness (r=0.449,P=0.047;r=0.487,P=0.030).In the diagnosis of HVPG≥12 mmHg,the area under curve (AUC) of liver stiffness was 0.875,the optimal cut-off value was 1.77 m/s,the sensitivity was 68.6 ％ and the specificity was 100.0％.In the diagnosis of HPVG≥20 mmHg,the AUC of liver stiffness was 0.798,the optimal cut off value was 1.85 m/s,the sensitivity was 100.0％ and the specificity was 68.8％.The AUC of spleen stiffness was 0.820,the optimal cut-off value was 3.23 m/s,the sensitivity was 100.0 ％ and the specificity was 56.3％.Conclusion In patients with liver cirrhosis,liver stiffness and spleen stiffness assessed by ARFI are positively correlated with HVPG and therefore ARFI has certain application value in the noninvasive diagnosis of portal hypertension.

To study the effect of endotoxin on liver apoptosis, L02 liver cells were cultured and passaged in vitro, and then stimulated by endotoxin at 10 mg/mL for 4, 8, 16 and 24 h respectively. Liver apoptosis was flow cytometrically and fluorescently detected. Immunohistochemistry was used to detect the delivery of smac and caspase9. The delivery of liver cell smac and the activity of caspase3 were measured by caspase3 assay kit. The hepatic failure models of rats were established by using D-galactosamine. The blood serum and liver tissues were collected for the detection of the liver function, the level of endotoxin and the activity of caspase3 by using chromogenic substrate limulus amebocyte lysate method (LAL) and caspase3 active assay kit. The expression of smac and caspase9 in liver cells was detected by Western blotting. With in vitro study, the L02 cells stimulated by LPS condensed into conglobation and formed apoptotic bodies. After those cells were stained by hoechst, the apoptotic cells displayed blue color under the fluorescent microscope. The apoptosis rate was increased over time and the apoptosis was mainly of advanced stage. Meanwhile, the rate of smac delivery and activity of caspase9 and caspase3 were increased on L02 cell membrane. In vivo, hepatic failure and obvious endotoxemia were induced by injection of more than 200 mg/kg D-GalN. Hepatic mitochondria smac was reduced with dosage of D-GalN and, on the contrary, the activity of caspase3 was increased. D-GalN at 200 mg/kg increased Caspase9 while D-GalN at 300 mg/kg decreased caspase9. Mitochondria signal channel plays an important role in the endotoxin-induced apoptosis of hepatic cells by promoting the release of smac from mitochondria to cytoplasm and activating caspase9 and caspase3 in its low-level channel.
Apoptosis/*drug effects ; Carrier Proteins/*metabolism ; Caspase 3/metabolism ; Caspase 9/metabolism ; Cell Line ; Cells, Cultured ; Endotoxins/*pharmacology ; Liver/cytology ; Liver/*metabolism ; Liver/pathology ; Liver Failure/chemically induced ; Liver Failure/pathology ; Mitochondrial Proteins/*metabolism ; Rats, Wistar

Objective:To determine the effect of Chuju total flavonoids (CJTF) on the secreted frizzled-related protein 4 (SFRP4) expression in Wnt pathway in rheumatoid arthritis (AR) model rats. Methods:hTe role of CJTF in the treatment of AR model rats was evaluated by rat arthritis score and paw edema score. The expression regulation of the SFRP4,β-catenin and C-myc in Wnt pathway in AR model rats was detected by RT-PCR and Western blot atfer CJTF gavage treatment. Results:Atfer CJTF treatment, the rat arthritis score and paw edema score in AR model rats were signiifcantly decreased when the AR model rats were treated with CJTF, the SFRP4 expression was signiifcantly up-regulated, while theβ-catenin and C-myc gene expression were signiifcantly down-regulated in AR model rat synovial tissues. Conclusion:CJTF has significant therapeutic effect and inhibitory effect on Wnt pathway activation by targeting SFRP4 in AR model rat synovium.

BACKGROUND: It is a hot investigation to many scholars that how to cure and prevent renal ischemic reperfusion injury in a utility way, but the mechanism is unclear at present. The investigation indicates that aquaporiin-1 plays an important role during this process. OBJECTIVE: To research the correlation between aquaporin-1 expression and renal function change following renal ischemic reperfusion injury. DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed at Histology and Embryology Laboratory of Dalian Medical University from June 2007 to December 2008. MATERIALS: A total of 80 healthy female adult Wister rats were randomly divided into control group and ischemia-reperfusion group. Rats in each group were observed at days 1, 2, 3, 5, and 7 after operation, with 8 rats for each group. The ischemia-reperfusion injury was established on the left kidney. METHODS: Right kidney was removed. The left renal pedicle was freed and occlused to establish ischemia-reperfusion injury model. After 40 minutes, the blood was re-flowed. If the kidney colored from dark red to bright red within 2-5 minutes, the ischemia-reperfusion injury models were successfully established, and the thrombus was not formed in the kidney vessels. If the kidney was still dark red after 5 minutes, the thrombus was formed, and the rats were excluded from the ischemia-reperfusion group. The abdomen was sutured after 40 minutes.MAIN OUTCOME MEASURES: Rats were sacrificed at days 1, 2, 3, 5, and 7 after ischemia-reperfusion injury. Samples of urine, serum, and kidney were performed with the examinations of urine, renal function, renal pathology and morphology, immunohistological assay of aquaporiin-1, and RT-PCR assay. RESULTS: After ischemia-reperfusion injury, the rats had hydrouria, urine osmotic pressure depress, symptoms of carnine and urea nitrogen increasing. HE staining demonstrated that renal tubular epithelial cells were swelling, necrosis, and desquamate. Aquaporin-1 expression and its mRNA level was decreased; in particular, the expression and level were the lowest at day 1 after ischemia-reperfusion injury and recovered to normal value at day 5 after ischemia-reperfusion injury. CONCLUNSION: The down expression of aquaporin-1 maybe one of the important indicators to reflect renal functional changes of acute renal failure following renal ischemia-reperfusion injury.