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OBJECTIVE To investigate the effects of astilbin （AST） on myocardial ischemia-reperfusion injury （MIRI） in rats and its potential mechanism. METHODS SD male rats were randomly divided into sham operation group， model group， positive control group （Compound Salvia miltiorrhiza tablets， 240 mg/kg）， AST low-dose and high-dose groups （30， 90 mg/kg）， and high- dose of AST+hypoxia-inducible factor-1α（HIF-1α） inhibitor group （AST 90 mg/kg+2ME2 15 mg/kg）， with 25 rats in each group. Except for sham operation group， MIRI model was induced in other groups， and then given relevant drug or normal saline intragastrically or intraperitoneally， for consecutive 28 d. Serum contents of cardiac troponin I （cTnI） and creatine kinase isoenzyme （CK-MB） were detected； volume ratio of myocardial infarction was measured； the pathological changes of myocardium， the apoptotic rate of myocardial cells and ultrastructure of mitochondria in myocardial tissue were all observed. The contents of tumor necrosis factor α （TNF-α）， interleukin 6 （IL-6） and malondialdehyde （MDA）， the activity of superoxide dismutase （SOD）， the expressions of HIF-1α， adenovirus E1B interacting protein 3 （BNIP3） and myosin-like Bcl-2 interacting protein （Beclin1） were determined in myocardium. The ratio of microtubule-associated protein light chain 3 （LC3） Ⅱ to Ⅰ （LC3 Ⅱ/Ⅰ） in rat myocardium was calculated. RESULTS Compared with model group， no obvious swelling was found in the myocardial tissue of rats in positive control group， AST low-dose and high-dose groups， and the myocardial fibers were arranged regularly； the volume ratio of myocardial infarction， the contents of cTnI， CK-MB， TNF-α， IL-6 and MDA， the apoptotic rate were decreased significantly （P＜0.05）， while SOD activity， protein expressions of HIF-1α， BNIP3 and Beclin1， LC3Ⅱ/Ⅰ were increased significantly （P＜0.05）. HIF-1α inhibitor could significantly weaken the improvement effect of AST on the above indicators in MIRI model rats （P＜0.05）. CONCLUSIONS AST enhances mitochondrial autophagy by activating HIF-1α/BNIP3 signaling pathway， thereby reducing MIRI in rats.
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Objective:To investigate eye movement desensitization and reprocessing on ICU memory and post-traumatic stress disorder among patients with ventilator associated pneumonia.Methods:Totally, 66 cases of ICU patients with ventilator associated pneumonia enrolled at Guangzhou Red Cross Hospital from August 2017 to August 2020 were collected as research object, they were randomly divided into intervention group and control group, with 33 cases in each groups. The control group received routine nursing care and cognitive training, in the intervention group, patients implemented EMDR therapy for 8 times. Before and after intervention, the effect was assessed by ICU memory scale and PTSD Checklist-Civilian Version, respectively.Results:After intervention, the scores of factual memories, emotional memories, delusional memories dimension and total ICUMS were (19.48±5.14), (10.45±2.62), (7.06±2.62) and (37.00±6.12) in the intervention group, those scores in the control group were (24.58±7.38), (13.58±4.07), (9.48±3.57) and (47.64±9.99), the difference was statically significant (t values were 3.14-5.22, all P<0.05). The re-experiencing, avoidance, hyperarousal dimensions scores and total PCL-C scores were (8.09±2.13), (10.42±2.62), (7.24±2.69) and (25.76±4.25) in the intervention group, and in the control group those scores were (9.30±2.35), (12.73±2.11), (9.24±2.51) and (31.27±4.97), the difference was statically significant (t values were 2.20-4.85, all P<0.05). Conclusions:Eeye movement desensitization and reprocessing therapy can effectively alleviate ICU memory and improve PTSD symptoms of patients with ventilator associated pneumonia.
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Objective:To observe the effect of emotional resilience training on compassion fatigue and psychological resilience of intensive care unit nurses.Methods:A total of 70 intensive care unit nurses were randomly divided into experiment group and control group, each of 35 cases. Control group nurses were given routine psychological guidance intervention, in addition, the experiment group was carried out 8 weeks of emotional resilience training. The effect of intervention was assessed by Chinese version of the compassion fatigue scale and Connor-Davidson resilience scale before and after intervention.Results:After intervention, the average scores of compassion satisfaction, secondary traumatic stress and total compassion fatigue scores were (26.46±4.99)pionts, (17.80±4.05)pionts and (69.14±5.70)pionts, significantly lower than in the control group (28.86±4.14)pionts, (20.11±4.38)pionts and (75.51±10.32)pionts, the difference was statistically significant ( t value was 2.190, 2.296, 3.197, P<0.05). After intervention, the average scores of tenacity, power, optimism and total resilience scores were (34.23±6.06)pionts, (25.77±5.01)pionts, (14.31±3.22)pionts and (74.31±9.55)pionts, significantly higher than in the control group (30.86±6.23)pionts, (23.31±3.29)pionts, (12.11±2.04)pionts and (66.29±7.28)pionts, the difference was statistically significant ( t value was 2.295-3.956, P<0.05). Conclusion:Emotional resilience training can effectively reduce the compassion fatigue and improve psychological resilience of intensive care unit nurses.
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Objective To investigate the ABO blood type and sepsis complicated with acute kidney injury,to provide the basis for prevention and treatment.Methods A total of 130 patients with sepsis from renmin hospital of Wuhan University intensive care unit from january 2015 to december 2015 were enrolled in this study,divided into complicated AKI group (64 patients in the observation group) and non-complicated AKI group (66 patients in the control group),analyzed two groups of general data,laboratory indicators,multivariate logistic regression analysis was used to screen out the risk factors for AKI in patients with sepsis.Results A total of 64 patients with AKI were collected from the observation group and 66 patients with non-AKI in the control group,the age of the observation group was higher than that of the control group (65.7 ± 13.1 years old vs 58.5 ± 15.4 years old,P =0.005),male proportion was higher than control group (76.6％ vs 56.1％,P =0.014),drop calcium pigment original quantitative was higher than control group (28.1 ± 21.0pg/L vs 21.1 ± 13.61μg/L,P =0.026),positive blood culture rate was higher than in the control group (30.2％ vs 15.3％,P =0.006).There was no significant difference in ABO blood group distribution between the two groups (P =0.825).The levels of white blood cell count,C-reactive protein and partially activated thrombin were higher in the observation group than in the control group,the platelet count and albumin level were lower than those in the control group,the difference was not statistically significant.The risk factors associated with the incidence of sepsis with AKI were analyzed by multivariate analysis of the logistic regression model:age(P =0.021,OR =0.965),gender (P =0.003,OR =5.321),calcitonin-original(P =0.047,OR =0.975),positive blood culture (P =0.002,OR =1.009),comparison of type A blood and type O blood (P =0.037,OR =5.409) were associated with sepsis complicated with AKI.Conlusion Type A blood and sepsis with AKI associated with the existence of independent correlation,type A blood may increase the risk of sepsis with AKI.
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Objective To observe the effect of costimulatory molecule B7-1 on cytoskeleton rearrangement in mouse podocytes induced by angiotensin Ⅱ (Ang Ⅱ),and to study the underlying molecular mechanism of B7-1 in the pathological changes of podocytes.Methods All cultivation of conditionally immortalized mouse podocytes (MPC) in vitro were divided into the following groups:normal control group,CTLA-4 group,Ang Ⅱ group (10-6 mmol/L 12 h,24 h;10-8 mmol/L 12 h,24 h) and CTLA-4 with Ang Ⅱ group.Transfect B7-1 RNA interference fragment (siRNA) to the mature podocytes,and then restimulated by Ang Ⅱ (10-6 mmol/L 12 h),the change of podocyte cytoskeleton after Ang Ⅱ stimulation were observed.The expression of B7-1 in each group was assayed by flow cytometry and Western blotting.The nephrin and p-nephrin protein levels in the four groups were also analyzed by Western blotting.At the same time,the podocyte cytoskeleton distribution as indicated by F-actin was observed by fluorescence microscopy.Results Flow cytometry and Western blotting showed that B7-1 was not expressed in the normal control group.Ang Ⅱ showed a concentration and time dependent induction of B7-1 expression in mouse podocytes (P ＜ 0.05).Western blotting indicated that Ang Ⅱ induced B7-1 protein expression (P ＜ 0.05).Expression of nephrin and p-nephrin was significantly down-regulated by Ang Ⅱ (P ＜ 0.05).Compared with the normal control group,the expression of podocyte protein nephrin and p-nephrin in Ang Ⅱ stimulation group was significantly reduced (P ＜ 0.05).Using FITC phalloidin fluorescence staining showed that CTLA-4+Ang Ⅱ stimulation group cytoskeleton rearrangement was improved significantly and F-actin recombinant score (mCFS) decreased compared with Ang Ⅱ group (P ＜ 0.05),suggesting that Ang Ⅱ led to the disorder of the podocytes cytoskeleton and the destruction of the cytoskeleton of podocytes by Ang Ⅱ could be improved after B7-1 blocking.Compared with the Ang Ⅱ stimulation,transfection of B7-1 siRNA + Ang Ⅱ stimulation group improved F-actin cytoskeletal rearrangement,and mCFS also decreased significantly (P ＜ 0.05),suggesting that transfection of B7-1 siRNA might improve the damage of Ang Ⅱ on podocytes cytoskeleton.Conclusions B7-1 participates in the process of cytoskeleton reconstruction and plays an important role in the pathological changes of podocytes.
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Objective To investigate the effects of nicardipine on apoptosis in ischemia?reperfusion myocardium of rabbits. Methods Forty New Zealand rabbit were randomly divided into 4 groups:control group(group A),ischemia group(group B),ischemia?reperfusion group(group C),ni?cardipine treatment group(group D). Ischemia?reperfusion model was established by left circumflex branch coronary artery ligation for 40 min,fol?lowed by 120 min reperfusion. Apoptotic cardiomyocyte was detected using the analysis for the exposure of phosphatidyl?serine ( Annexin V)meth?od. Results Myocardial cell damage was lower in D group than that in group B and group C from pathology(P0.05). Conclusion Nicardipine horizon can reduce the myocardial cell apoptosis in rabbits with myocardial ischemic reperfusion injury.
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Objective To investigate the effect of on serum and urine MCP -1 secretion in patients with Stage IV Type 2 diabetic nephropathy treated by lipoic acid .Methods We enrolled 76 diabetic nephropathy patients ,who were randomly divided into two groups . Patients in group T (24 males and 14 females) were treated by lipoic acid 0.6 gram per day.Those in group C (22 males and 16 fe-males) were treated by routine drugs.The serum creatinine (Scr), urea nitrogen (BUN), fasting blood sugar(FBS) and 24h urinary pro-tein,serum and urine MCP -1 secretion were measured at the experiment onset and 3 weeks later.Results There was no significant difference in the serum creatinine , urea nitrogen, fasting blood sugar, HbA1c between the two groups neither at the experiment onset nor after 3 weeks.Compared to experiment onset , 24h urine protein (Tp/24h), serum and urine MCP-1 secretion were all significantly de-creased (P<0.05) in group T after 3 weeks.Compared to group C (2.41 ±0.91g/24h, 91.45 ±33.41pg/ml, 114.78 ±36.35pg/ml), all the levers in group T (1.89 ±0.72g/24h, 39.50 ±13.68pg/ml, 63.41 ±19.57pg/ml) was significantly decreased (P<0.05) after 3 weeks.There was a positive correlation between the serum , urine MCP-1 levels and Tp/24h (r=0.572, P<0.05;r=0.697,P<0.05).Conclusion Lipoic acid can reduce urine protein excretion in diabetic nephropathy patients , maybe by decreasing serum and u-rine MCP-1 secretion .
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Objective To investigate association between serum uric acid (SUA),albuminuria and glomerular filtration rates (eGFR) in type 2 diabetic patients.Methods A total of 220 patients were enrolled in this cross-sectional study.According to urinary albumin excretion rates,patients were divided into 3 groups:normoalbuminuria (NAU) group,microalbuminuria (MAU) group,and macroalbumnuria group (MAAU).The first two groups were subdivided at SUA ＞ 420 μmol/L (＞ 357 μmol/L,female) into normouricemia group and hyperuricemia group,at eGFR ＞ 90 ml/min into high and low renal function groups.General information,blood biochemical results were collected to analyze the association between serum uric acid,eGFR,UAER and urine albumin quantification among different groups.Results The difference of SBP,duration of diabetes (DD),Scr,SUA and eGFR between every two groups were significant (P ＜ 0.05).SBP,DD,Scr and SUA were highest in subjects with macroalbumnuria,second in microalbuminuria group,and lowest in normoalbuminuria group,while eGFR was lowest in macroalbumnuria group and highest in normoalbuminuria group.Prevalence of hyperuricemia in macroalbumnuria group (56.9％) and microalbuminuria group (51.2％) were also significantly higher than that in normoalbuminuria group (17.5％) (all P ＜ 0.01).The difference of UAER in the subgroups of normouricemia and hyperuricemia was more significant in microalbuminuria group than in normoalbuminuria group.eGFR was significantly lower in hyperuricemia subgroups (P ＜0.01).Age and SUA were significantlg higher in subjects with low renal function compared with high eGFR (P ＜ 0.05).Linear regression analysis indicated SUA was negatively correlated with eGFR after adjusted age,DD and UAER (β =-0.430,P ＜ 0.01).Binary logistic regression analysis found that increased age,DD and SUA were risk factors of microalbuminuria β =1.092,95％ CI(1.025,1.163),P ＜ 0.01;β =1.005,95％CI(1.001,1.009),P ＜ 0.05;β =1.407,95％ CI(1.052,1.881),P ＜ 0.05)] and SUA,age were risk factors of early renal function decline [β =1.015,95 ％ CI(1.00,1.023),P ＜ 0.01;β =1.098,95％ CI(1.006,1.199),P ＜ 0.05].Conclusion SUA is independently associated with albumnuria and renal function decline in type 2 DM patients.
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Objective To investigate the role of IQ domain GTPase-activating protein 1 (IQGAP1) in angiotensin Ⅱ (Ang Ⅱ)-induced podocyte apoptosis and the underlying mechanism.Methods Differentiated mouse podocytes were exposed to Ang Ⅱ at different concentrations for 6 h or at 10-8 mol/L for variable incubation time.Podocyte apoptosis was assessed by flow cytometry.Expression of IQGAP1 was analyzed by immunofluorescence and Western blotting.IQGAP1 siRNA and MAPK pathway inhibitors(10 μmol/L SB202190,25 μmol/L SP600125,10 μmol/L U0126) were further introduced to investigate the role of IQGAP1 and MAPK signalings in the process.And coimmunoprecipitation was used to evaluate the interaction between ERK1/2 and IQGAP1.Results (1)Ang]] promoted podocyte apoptosis in a dose-and time-dependent manner.(2) IQGAP1 was located in celluar membrane and cytoplasm of cultured podocytes.Exposure to Ang Ⅱ stimulated IQGAP1expression in a dose-and time-dependent manner,and elevated phosphorylation of p38,JNK,and ERK1/2 simultaneously.(3) Pretreatment with SB202190,SP600125,or U0126 dramatically prevented Ang Ⅱ-promoted podocyte apoptosis respectively (P ＜ 0.05).However,the protein level of IQGAP1 was not altered.(4) Knockdown of IQGAP1 with siRNA obviously prevented Ang]Ⅱ-induced apoptosis of podocytes(P ＜ 0.05) and reduced Ang Ⅱ-induced phosphorylation of ERK1/2(P ＜ 0.05),but not that of p38,JNK.This was accompanied by a reduced interaction between ERK1/2 and IQGAP1(P ＜ 0.05).Conclusion IQGAP1 contributes to Ang Ⅱ-induced podocyte apoptosis by interacting with the ERK1/2 signaling protein.
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BACKGROUND:There are few clinical control ed trials about the clinical effects in patients with pertrochanteric femur fractures after treatment with short or long proximal femoral nail antirotation. OBJECTIVE:To compare the clinical outcomes in patients with AO/ASIF-A1/2 pertrochanteric femur fractures after treatment with short or long proximal femoral nail antirotation. METHODS:A total of 98 patients with AO/ASIF-A1/2 pertrochanteric femur fractures were treated by proximal femoral nail antirotation. They were divided into two groups according to the type of proximal femoral nail antirotation:short nail group (n=50) and long nail group (n=48). The operative time, blood loss, and hospital stay were recorded in both groups. In fol ow-up, fracture healing time, imaging and clinical complications were evaluated. In the final fol ow-up, Harris hip score was used to evaluate functional recovery. RESULTS AND CONCLUSION:Compared with the short nail group, operative time was shorter and blood loss was less in the long nail group (P0.05). Average fol ow-up periods were respectively (15.8±6.4) months and (16.2±5.7) months in the long nail and short nail groups.“Cutting-out”or infection occurred in five patients in the long nail group and three in the short nail group. Besides above-mentioned patients, the remaining patients in the two groups achieved fracture healing. No significant difference in average fracture healing time was detected between groups (P=0.588). In the final fol ow-up, no significant difference in Harris hip score was detectable in the two groups (P=0.204). The incidence rates of total postoperative complications in the long and short nail groups were 31.2%and 16.0%, respectively (P=0.075). Results suggested that no differences in the union and complication rates between the two groups were identified, suggesting that long nails offer no advantage compared with short nails for stabilizing AO/ASIF-A1/A2 pertrochanteric femur fractures.
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To investigate the efficacy, safety and cost of treating patients with acute kidney injury [AKI] and diabetic nephropathy [DN] with continuous renal replacement therapy [CRRT] or sustained low-efficiency daily diafiltration with hemofiltration [SLEDD-f]. Medical records of patients with AKI/DN from January 2006 to December 2012 were reviewed. Fifty-five patients who received CRRT and 52 who received SLEDD-f were included in the study. CRRT and SLEDD-f were performed for 20-72 h per session and 8-10 h per session, respectively. Mortality and renal function recovery rates were evaluated 30 days after the initiation of renal replacement therapy [RRT] and APACHE-II and SOFA scores, anticoagulant dose, inflammatory indices and cost were calculated at baseline and at the end of RRT. Of the 55 patients treated with CRRT, 49 [89.1%] had a 30-day survival rate and 30 [54.5%] had a 30-day renal recovery rate. Of the 52 patients with SLEDD-f, these rates were 92.3% [n = 48] and 61.5% [n = 32], respectively. The dosage of low-molecular-weight heparin in the CRRT and SLEDD-f groups was 15,230 +/- 1,460 and 6,320 +/- 490 U/day, respectively. The cost of hemopurification and the total cost for patients treated with CRRT was CNY 28,628 +/- 5,576 [USD 4,210 +/- 820] and CNY 38,828 +/- 6,324 [USD 5,710 +/- 930], respectively. These were higher than those for patients treated with SLEDD-f at CNY 13,260 +/- 1,564 [USD 1,950 +/- 230] and CNY 19,720 +/- 2,652 [USD 2,900 +/- 390], respectively. SLEDD-f offered a similar chance of renal recovery and also had further advantages such as a lower heparin dosage, a shorter therapy time and lower hospitalization costs for patients than CRRT. Studies with larger, randomized sample sizes are needed to confirm these findings
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Humans , Acute Kidney Injury , Renal Replacement Therapy , HemofiltrationABSTRACT
Objective To evaluate the effects of high glucose on autophagy and apoptosis of podocyte and explore the signaling pathway in high glucose-induce podocyte autophagy.Methods Differentiated mouse podocytes were exposed to high glucose(30 mmol/L) or rapamycin (autophagy enhancer,1 μg/L) or LY294002 (a selective PI3K inhibitor,50 mmol/L) for 24 h.The formations of autophagy were observed by electron microscopy and acridine orange staining.Apoptosis was evaluated by flow cytometry.The expression of autophagy protein LC3-Ⅱ/Ⅰ and Beclin-1 as well as the phosphorylation of AKT and mTOR were examined by Western blotting analysis.Results High glucose induced podocytes apoptosis,increased autophagy and the expression of autophagy-associated proteins (all P ＜ 0.05).Rapamycin further increased the expression of LC3-Ⅱ and Beclin-1 protein (all P ＜ 0.05),but LY294002 inhibited partialiy the protein expression of LC3-Ⅱ and Beclin-1 induced by high glucose (both P ＜ 0.05).Treatment with rapamycin increased the phosphorylation of AKT,but reduced that of mTOR in podocytes.Moreover,LY294002 inhibited phosphorylation of both AKT and mTOR (both P ＜ 0.05).Conclusions High glucose promotes podocyte autophagy and apoptosis.High glucose-induced autophagy is mediated partly through PI3K-AKT-mTOR signaling pathway.
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Objective To investigate the effects of angiotensin1-7 (Ang1-7) on renal tubulointerstitial fibrosis of diabetic rats.Methods Thirty-two male Wistar rats were randomly divided into four groups: normal control group,diabetic group,telmisartan group,Ang1-7-treated group.For 9 weeks after diabetes mellitus model established,24 h proteinuria,urine NAG/Cr,glucose,insulin,TG,TC,BUN,Scr,Na+ and K+ were assessed.Renal pathological changes were evaluated by PAS staining; Expression of TGF-β1,PPARγ and α-SMA mRNA was deteeted by real-time PCR; Protein levels of PPARγ,α-SMA and TGF-β1 were detected by Western blotting.Results (1)At the end of the ninth week,the blood pressure,proteinuria,renal weight/body weight in group DM were significantly higher than those in group NC (P＜0.05).(2)Renal interstitial fibrosis in group DM was obviously severe as compared to group NC (P＜0.05),but was improved in group TM and group T(P＜0.05).(3)TGF-β1 and α-SMA mRNA in group DM were significantly increased,and PPARγ mRNA was significantly decreased.Compared with group DM,TGF-β1 and α-SMA mRNA were significantly decreased,and PPARγ mRNA was significantly increased in group TM and group T,especially in group T.(4)TGF-β1 and α-SMA in group DM were significantly increased,and PPARγ decreased significantly.Compared with group DM,TGF-β1 and α-SMA decreased significantly,PPARγ increased significantly in group TM and group T,especially in group T.Conclusion Ang1-7 inhibits high glucose-induced α-SMA expression in vivo through up-regulating the PPAR expression and may inhibit renal tubulointerstitial fibrosis of diabetic rats.
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ObjectiveTo evaluate the effects of angiotensin Ⅱ (Ang Ⅱ )infusion on renal c-Abl expression in vivo,and on podocyte c-Abl expression change in cultured mouse podocytes.Methods Twenty four male Sprague-Dawley rats (Group C,D,E and F) were assigned to receive Ang Ⅱ(400 ng· kg-1 min-1) by osmotic minipump and of which 12 rats (Group D and F) were assigned to receive telmisartan (3 mg·kg-1·d-1),six rats received normal saline(Group B),and six rats were used as normal control(Group A).Animals were sacrificed at day 14 (Group C and D),day 28 (Group E and F) respectively.Conditionally immortalized mouse podocytes were used in vitro.Podocytes were studied 2 weeks after thermoswitching from 33℃ to 37℃.Cells were fetal bovine serum(FBS) starved for at least 12 hours prior to stimulation.The cultured podocytes were treated withAngⅡdosesranging from10 -9 mol/L to10 -6 mol/L andfor differenthours.Expression of renal and podocytes c-Abl was examined by immunofluorescence staining,real-time PCR and Western blotting.Results(1) Distribution of c-Abl expression was mainly in the cytoplasm and nuclear of the podocytes in vivo and in vitro. (2) Expressions of c-Abl mRNA and protein wereincreasedinAng Ⅱ-infusedratpodocytesandAng Ⅱ-inducedculturedmouse podocytes(P＜0.05),and the effects of Ang Ⅱ were dose-dependent and time-dependent in vitro.Conclusion There are c-Abl mRNA and protein expression in podocytes,and c-Abl may play a critical role in the pathogenesis of Ang Ⅱ -induced podocyte injury.
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Objective To investigate the effects of adhesion mediated by bone nlalTow stroma celh from leukemia patient on chemotherapeutics sensitivity and cell cycle of Jurkat cells in the co-cultured model.Methods Bone mw stroma cells were isohted and cultured from leukemia patients routinely.To construct the co-cultured model.Jurkat cells were co-cultured with BMSCs the irradiated layer by 60Co,and the model was observed with scanning electron microscope.The IC50 values of Jurkat cells expesured to DNR were quantified by MTT.The cell cycles of Jurkat cells after 24h-adhesion in the co-cultured model were analyzed by Facs.The expression of cyclin A,cyclin E and p27 in Jurkat cells adhered to BMSCs for 4h.24h and 48h were detected by Western blot.Results Jurkat ceUs in the co-cultured model showed a decreased sensitivity to DNR.ICSO values for normal BMSCs,leukemic BMSCs and non-adhered control were of 1.78Ixmol/L,2.30pznol/L and 0.45p,mol/L,respectively.The percentages of Go-Gl phase for leukemic BMSCs group and non-adhered control group were of 48.74％±8.77％and 27.83％壬1.86％.Respectively.The percentages of Gz-M phase for leukemic BMSCs group and non-adhered control group were of2.01％±1.17％and 20.33％±1.84％。Respectively.Compared with eomrol group,the 24h-ad- hesion mediated by BMSCs from leukemia patients up-regulated the percentage of Go-G1 phase of Jurkat cells(P<O.01),but down-regula-ted the percentage of G2-M phase(P<O.01).The expression of cyclin A and cyclin E in Jurkat ceUs was up-regulated when adhered to BMSCs for four hours and the higher expression emerged after adhering for 24h and 48h.Oppositely,the expression of p27 were down-regulated.Especially after 48h-adhesion.Conclusions Drug resistance of leukemia cells csn be mediated by adhering to bone marrow stroma cells,which may be related to the changes of cell cycle.
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Objective To investigate the chemotherapeaties sensitivity and expression of bcl-2, bcl-xL and bax of Jurkat cells co-cultured with bone marrow stromal cells (BMSC) isolated and cultured from leukemia patients. Methods BMSC were isolated and cultured from leukemia patients routinely. To construct the co-cultured model, Jurkat cells were co-cultured with of the irradiated layer BMSC by 60Co and observed the model with scanning electron microscope. The Jurkat cells suspension-cultured were used as control. The apoptosis and IC50 were detected by the FACS can machine and MTT, respectively. The expression of bcl-2,bcl-xL and bax in Jurkat cells was detected by Western blotting. Results We found that the Jurkat cells in the model showed a decreased sensitivity to DNR, IC50 values for leukemic BMSC and nonadhered contol were of 2.30 μmol/L and 0.45 μmol/L, respectively. Moreover, Jurkat cells adhered to BMSC have a survival advantage over suspended cells following DNR exposure for 24 h, apoptosis percentages for leukemic BMSC group and nonadhered controls were of (6.05±0.54)% and (25.74±6.15)%, respectively. As compared with controls, leukemic BMSC group had significant difference in apoptosis percentages (P <0.01). The expression of bcl-2 in Jurkat cells was up-regnlated when adhered to BMSC for 4 h and the higher expression emerged after adhering for 24 h and 48 h. No marked change of bcl-xL and bax expressions were observed in the adhered Jurkat cells. Conclusion The adhered-culture with bone marrow stromal cells isolated from leukemia patients could make the leukemia cells acquire drug resistance, which was associated with the up-regulated expression of bcl-2 in the leukemia cells.
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Objective To explore the feasibihty of TRAIL to be used to remove the leukemia cells from the autologous hemopoietic stem cell transplants. Methods The expression of decoy receptor 1 and decoy receptor 2 on the bone marrow mononuclear cell were routine-ly isolated and observed by fluorescence microscope after PE-DcR1 or PE-DcR2 stain. The apeptosis rates of mononuclearcell and Jurkat cells interfered by 200ng/ml TRAIL for 18h were determined by flow cytometry after AnnexinV/Pl stain. The interfered mononuclear cells were cultured to count the number to form colony-forming unit-fibroblast(CFU-F). The Jurkat cells labeled by green fluorescent protein were in-corporated into the mononuclear ceils and affected by 200ng/ml TRAIL for 24h. The incorporated cells were cultured in the system with GM-CSF and EPO and the numbers of the CFU-GM, BFU-E and fluorescence colony were counted on the seventh day. Results Both decoy re-ceptor land decoy receptor 2 of TRAIL can be detected on membrane or in cytoplasm of the bone marrow mononuclear cell. The apoptosis rate of mononuclear cell interfered by 200ng/ml TRAIL for 18h was (5.95±1.23)%, which was markedly lower than that of Jurkat cells (33.42±2.28) %. The number of CFU-F of TRAIL group and control group were 235.67 ~ 33.56 and 249.33±42.72, respectively. No marked difference can be found between the mentioned two groups. Moreover, TRAIL decreased the number of fluorescence colony formed by Jurkat cells without significant decreasing the number of CFU-GM and BFU-E formed by bone marrow mononuclear cells. Conclusion TRAIL can selectively induce apoptosis in Jurkat cells without marked toxic effect on the bone marrow mononuclear cells, which means that TRAIL can be used to remove the leukemia cells from the autologous hemopeietic stem cell transplants in vitro.
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Objective To clarify the risk,treatment and preventive measurement of transfusion related acute lung injury (TRALI) occurred in HLA half-matched hematopoietic stem cell transplantation. Methods A case of TRALI occurred in HLA half-matched hematopoietic stem cell transplantation was analyzed,including the clinical characteristics,the laboratory and instrumental examination,the treatment measure and prognosis,and then the associated literature was reviewed.Results Owing to the blood products(fresh platelet) transfusion during transplantation,the patient got TRALI. And after active rescue,the patient eventually died due to the worsen condition.Conclusion The risk of TRALI is very high in HLA half-matched hematopoietic stem cell transplantation since the blood products transfusion is inevitable,so the effective and timely treatment and preventive measurement are necessary.
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BACKGROUND: Increase of fibrinogen is an independent risk factor for cardio-cerebrovascular disease and peripheral angiopathy. Fibrinogen and its degradation product exist in carotid atheromatous plaque and it can stimulate proliferation and migration of smooth muscle cells so as to play a key role in early atherosclerosis.OBJECTIVE: To analyze the relationship between plasma fibrinogen and carotid atheromatous plaque.DESIGN: Descriptive observation.SETTING: Department of Neurology, Xinhua Hospital, Medical Collegeof Shanghai Jiaotong University.PARTICIPANTS: A total of 81 patients with ischemic stroke were selected from Neurological Department of Xinhua Hospital affiliated to Medical College of Shanghai Jiaotong University from September to December 2001.There were 53 males and 28 females aged 41-85 years with the mean age of (65±11) years.METHODS: On the basis of carotid ultrasonography, 81 patients with ischemic stroke were divided into present plaque group (n=45) and absent plaque group (n=36) to measure plasma fibrinogen, serum total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and other risk factors of atherosclerosis so as to observe the correlation between risk indexes and carotid atheromatous plaque.MAIN OUTCOME MEASURES: Risk indexes of carotid atherosclerosis and carotid atheromatous plaque.RESULTS: All 81 patients were involved in the final analysis. ① Values of fibrinogen, TC and LDL-C in present plaque group were (4.38±1.33) g/L,(5.19±1.27) mmol/L and (3.15±0.37) mmol/L, respectively, which were higher than those in absent plaque group [(3.20±1.30) g/L, (4.56±1.30) mmol/L,(2.49±0.92) mmol/L, P ＜ 0.05]. ② Ratios of carotid atheromatous plaque were 11% (5 cases), 16% (7 cases) and 73% (33 cases) in fibrinogen groups at Iow, moderate and high dosages, respectively. There was significant difference (P =0.002). ③ Multiple factor analyses of carotid atherosclerosis showed that the risk factors which promoted the formation of carotid atheromatous plaque were diabetes mellitus, low density lipoprotein (LDL), fibrinogen and age growing.CONCLUSION: Fibrinogen plays an active role in development and progression of carotid atheromatous plaque.
ABSTRACT
Objective To study the anti-depressive mechanisms of acupuncture by investigating the effects of acupuncture on depressive behaviors and amino-neurotransmitter in hippocampus in rats treated with chronic mild unpredicted stresses.Methods Thirty adult SD rats were randomly divided into a normal group,a depressive model group and an acupuncture group,with 10 rats in each group.Separated feeding,long-term unpredictable and middle stimulation stress were used for development of depression rat model.At the same time,the third group was treated by acupuncture.Autonomic behavior was measured with Open Field test and concentration of amino-neurotransmitter in hippocampus was measured by high performance liquid chromatography coupled with fluorescence detection.Results Compared with the normal group,autonomic activity of the rats and the concentration of GABA were significantly reduced in the model group,while the concentration of Glu was significantly increased(P0.05).Conclusion Acupuncture could reverse the depressive behaviors of rats caused by middle stimulation stress,and its anti-depressive mechanisms is related to the function of regulating the concentration of amino-neurotransmitter in hippocampus.