1.The Effect of immunomodulatory therapy On CD4+CD25+ tregs in peripheral blood of patients with Sepsis
Fangqiang SONG ; Shifang DING ; Yongqin ZHOU ; Ming GUO ; Ming LI
Chinese Journal of Emergency Medicine 2011;20(3):249-252
Objective To investigate the percentage of CD4 + C125 +Tregs in peripheral blood of patients with sepsis and its effect on cell immunity so as to unravel the effect of immunomodulatory therapy on it. Method Fourty patients with sepsis in ICU were randomly (random number) divided into experimental group and control group . The patients of experimental group were treated with Ulinastatin and immunoregulation agent (Thymosin αl) as well. The blood specimens were collected just before treatment, 3 days and 8 days after treatment. The percentages of CD4 + CD25 + Tregs and lymphocyte subsets were detected by using FCM (flow cytometry), and TNF-α, IL-6 and IL-10 assayed by using ELISA, and APACHE Ⅱ scores were calculated. Results Before treatment, the percentage of CD4 + CD25 + Tregs increased, and the number of lymphocytes and the percentage of T lymphocytes decreased, especially the CD4 + T lymphocytes and CD4+/CD8+ decreased more markedly, and the levels of IL-6 and TNF-α increased. After treatment,the percentage of CD4+ CD25 + Tregs was decreased, the number of lymphocytes and CD4 +/CD8 + increased, and the levels of APACHE Ⅱ score, IL-6 and TNF-α decreased especially in the experimental group decreased more significantly (P < 0. 05). Conclusions The percentage of CD4 + CD25+ Tregs in peripheral blood can reflect the immune status of patients with sepsis and become a novel indicator to estimate the progress of sepsis, and the immunity and prognosis of patients. Treating the patients with Thymosin αl and Ulinastatin can raise their immunity, decrease the levels of IL-6, TNF-α and APACHE Ⅱ score and improve their prognosis.
2.Mechanism of Aldehyde Dehydrogenase-2 Regulated Human Endothelial Progenitor Cells Oxidative Stress Reaction
Qijun JIANG ; Zhigang GONG ; Zhigang LI ; Shifang DING
Chinese Circulation Journal 2016;31(5):502-507
Objective: To investigate the role of aldehyde dehydrogenase-2 (ALDH-2) for regulating human endothelial progenitor cells (EPCs) oxidative stress reaction and its mechanism. Methods: Human EPCs were isolated from peripheral blood of healthy adults and the cells were cultured in 4 groups:①Blank control group,②Alda-1 group, the cells were treated by 1μmol/L Alda-1, a speciifc activator of ALDH-2,③tBHP (10μg/ml) group and④Alda-1 pretreatment+tBHP group. EPCs reactive oxygen species (ROS) levels were evaluated by DCFH-DA staining, mitochondrial membrane potentials were detected by JC-1 method, migration capacity was measured by transwell chamber method and the activation of p38 signal pathway was examined by Western blot analysis. Results: Compared with Blank control group, ROS levels in tBHP group and Alda-1 pretreatment+tBHP group were (441.7 ± 24.8) % and (237.4 ± 12.0) %, allP<0.05. In Blank control group, tBHP group and Alda-1 pretreatment+tBHP group, the proportion of EPCs lost their mitochondrial membrane potentials were (5.7 ± 2.1) %, (81.7 ± 3.7) % and (37.4 ± 3.2) % respectively, allP<0.05; the number of EPCs migration were (108 ± 9)/HP, (22 ± 4)/HP and (67 ± 7)/HP respectively, allP<0.05. Compared with Blank control group, the activation of p38 signal pathway increased to (259.1 ± 7.7) % in tBHP group, while it was reduced to (186.4 ± 8.0) % in Alda-1 pretreatment+tBHP group. Conclusion: ALDH-2 could reduce ROS level in human EPCs, it may decrease mitochondrial membrane damage, protect migration which might be related to p38 signal pathway.
3.Akt-eNOS signal pathway for mediating norepinephrine regulating mobilization of endothelial progenitor cells
Qijun JIANG ; Zhigang GONG ; Zhigang LI ; Shifang DING
Chongqing Medicine 2016;45(19):2602-2605
Objective To investigate the effect of norepinephrine (NE) on the proliferation and migration capacity of endo‐thelial progenitor cells (EPCs) ,and bone marrow mobilization and to analyze its molecular mechanism .Methods The 8‐week old C57 mice were taken and randomly divided into 3 groups ,5 cases in each group :the blank control group(subcutaneous injection of normal saline without operaion) ,model group(subcutaneous injection of normal saline and ischemia in left lower extremity ) and NE group(subcutaneous injection of NE 100μmol/100 μL and ischemia in left lower extremity) .The limb ischemia model was prepared by adopting the femoral arterial ligation in mouse left lower extremity ,then NE was continuously pumped by the micro‐osmotic pump .The EPCs contents from bone marrow ,peripheral blood and spleen were assayed with the flow cytometric analyzer ;human peripheral blood EPCs were cultured and stimulated by NE .The proliferation and migration capacity ,and the activation situation of Akt and eNOS signal pathway were detected .Results NE could promote the mobilization of bone marrow EPCs in limb ischemia mice ,increased the EPCs quantity of peripheral blood and spleen ,comparing the NE group with the model group ,the EPCs quantity was increased for bone marrow [(3 .271 ± 0 .772)% vs .(1 .320 ± 0 .256)% ] ,peripheral circulation[(0 .261 ± 0 .041)% vs .(0 .110 ± 0 .028)% ] and spleen[(4 .671 ± 0 .345)% vs .(1 .880 ± 0 .0 .381)% ] ,the differences were statistically significant (P<0 .01) .NE could promote the proliferation and migration capacity ,moreover could activate the Akt‐eNOS signal pathway in EPCs with a dose dependent manner .Conclusion NE could promote the proliferation and migration of EPCs and mouse bone marrow mobilization via the Akt‐eNOS signal pathway .
4.Astaxanthin Inhibits Endothelial Progenitor Cell Apoptosis Induced by Oxidative Stress via Mitochondria-targeted Protective Mechanism
Zhigang GONG ; Shifang DING ; Qijun JIANG ; Wenbo FU
Herald of Medicine 2014;(6):712-717
Objective To investigate the effect of astaxanthin( ASX)on endothelial progenitor cells( EPCs)injury induced by oxidative stress in vitro and to explore its underlying mechanism. Methods Cultured EPCs isolated from peripheral blood were randomly divided into 5 groups:normal control,model group[ tert-butyl hydroperoxide( tBHP)100μmol·L-1 ],and ASX+tBHPgroups(thecellswerepreconditionedwithASX0.1,1.0,and10.0nmol·L-1,respectively).Thecellviabilitywas measured by MTT method. The level of reactive oxygen species( ROS)was determined by DCFH-DA method. The changes of mitochondrial membrane potential( MMP)and apoptosis ratio were detected by JC-1 method and DAPI method,respectively. caspase-3 activity changes of EPCs were detected. Results The cell viability of EPCs was improved with the increasing concentration of ASX. Compared with the model group[(48. 5±4. 3)%],0. 1,1. 0,10. 0 nmol·L-1 ASX significantly increased the cell viabilities[(57. 6±8. 2)%,(77. 6±7. 5)%,and(85. 3±6. 1)%,P﹤0. 05]. The results of DAPI staining revealed that ASX pretreatment could significantly reduce the apoptotic rate of EPCs. The apoptotic rate of the model group was( 27. 8 ± 3. 2)%,while that of ASX+tBHP groups was[(20. 4±2. 9)%,(14. 9±1. 7)%,and(7. 8±0. 7)%,P﹤0. 05],respectively. The data from caspase-3 activity assay indicated that ASX precondition could also remarkably decrease the caspase-3 activity for EPCs. The caspase-3 activity of the model group was(0. 345±0. 018),while that of the ASX+tBHP group were[(0. 291± 0. 013),(0. 209±0. 004),and(0. 169±0. 013),P﹤0. 05],respectively. In addition,treatment with tBHP resulted in an increase of DCF fluorescence,while ASX precondition could decrease the DCF fluorescence,which suggested the accumulation of intercellular ROS for EPCs. Injury of michondrial membrane resulted in the loss of mitochondrial membrane potential( MMP). The MMP detected by JC-1 method revealed that compared with model group,pretreatment of ASX inversed the reduction of MMP. Conclusion Astaxanthin inhibits endothelial progenitor cell apoptosis induced by oxidative stress through inhibiting ROS production,improving the mitochondrial function and down-regulating caspase-3 activity.
5.Analysis of the effect and safety of transradial percutaneous coronary intervention
Feng LIN ; Shifang DING ; Jiequan JIANG ; Zhinan CHEN
Chinese Journal of cardiovascular Rehabilitation Medicine 2015;24(4):425-427
Objective:To study the effect and safety of transradial percutaneous coronary intervention (PCI).Meth-ods:Clinical data of 306 patients,who received PCI in our hospital from Mar 2012 to Jan 2014,were retrospectively analyzed,including radial group (n=153),and femoral group (n=153).Therapeutic effect,time and postoperative complications etc.were compared between two groups.Results:A total of 151 cases completed PCI successfully in radial group,the success rate was 98.7%;a total of 150 cases completed PCI successfully in femoral group,the suc-cess rate was 98.0%,there was no significant difference in success rate of operation between two groups,P >0.05. Compared with femoral group,there were significant reductions in hospitalization time [(8.0±3.5)d vs.(3.5± 1.7)d],treatment cost [(3.74±2.06) × 104 yuan vs.(2.61 ± 1.4) × 104 yuan],P <0.01 both,in incidence rates of postoperative coronary occlusion (3.92% vs.0%),arrhythmia (11.76% vs.1.31%),vascular spasm (6.54% vs.1.96%)and hematoma (7.19% vs.0.65%)etc.in radial group,P < 0.05 or < 0.01. Conclusion:Transradial PCI possesses better effect than that of transfemoral ,and it can reduce hospitalization time,cost and postoperative complications,which is worth extending.
6.Influence of long-term oral valsartan on ventricular arrhythmia after myocardial infarction in rabbits
Zhinan CHEN ; Shifang DING ; Zhigang GONG ; Qing LU
Chinese Journal of cardiovascular Rehabilitation Medicine 2014;23(2):165-170
Objective: To explore influence of long-term oral valsartan-angiotensin II type 1 receptor blocker on ventricular arrhythmia after myocardial infarction (MI) in rabbits and its possible mechanism. Methods: A total of 24 New Zealand rabbits were randomly divided into sham operation group (n=8), MI group (n=8) and valsartan group (n=8) according to number table. Sham operation group only received thoracotomy without ligation of anterior descending branch of left coronary artery (LAD), while MI group and valsartan group received ligation of anterior descending branch of LAD. Valsartan group received valsartan gavage (10 mg•kg-1•d-1) since the second day after operation, three groups all were fed for 12 weeks. Mono active potential (MAP) of left ventricular myocardial cells of subendocardial myocardium(inner layer myocardium), subepicardial myocardium(outer layer myocardium)and middle layer myocardium were recorded before MI and 12 weeks after MI, and times of provocative malignant arrhythmias were recorded on 12 weeks after MI in three groups. Results: 1. Ventricular tachycardia or fibrillation (VT/ VF) episodes were markedly decreased in VAL group than that in MI group on 12 weeks after MI [(3.1±0.8) vs. (12.7±1.5), P<0.05]; 2. After MI 12 w, the action potential duration to 90% repolarization (APD90) of three-layer ventricular myocytes in MI group was prolonged than that before MI [(259.2±22.1)ms,(288.0±25.8)ms,(244.6±22.6)ms vs.(230.1±23.2)ms,(244.2±23.4)ms,(229.0±21.7)ms, P<0.05 or<0.01];but there were no significant difference in APD90 of three layers ventricular myocytes between before and after MI in valsartan group (P>0.05 all); Compared with sham operation group and valsartan group, there was significant prolonged in transmural dispersion of repolarization (TDR) [(18.8±6.2) vs. (23.9±7.7) vs. (37.2±10.2), P<0.05 or<0.01] in MI group; There was no significant difference in TDR between valsartan group and sham operation group (P>0.05). Conclusions: Long-term oral valsartan can significantly reduce malignant ventricular arrhythmia incidence in rabbits after MI, which may be related to improving TDR in rabbits after MI.
7.Variation in serum visfatin levels 24 hours after coronary stent implantation
Yun HOU ; Shifang DING ; Huijie WANG ; Juquan JIANG ; Wenbo FU ; Hua WANG ; Zhinan CHEN
Chinese Journal of Tissue Engineering Research 2014;(16):2619-2624
BACKGROUND:Coronary stent implantation can cause blood vessel damage and wal reconstruction, leading to vascular stent restenosis. Studies have found that visfatin is associated with inflammatory reaction, and exhibits an increased expression at the site of plaque rupture in acute myocardial infarction. OBJECTIVE:To investigate the influence of percutaneous coronary intervention on the levels of visfatin in patients with coronary heart disease. METHODS:Thirty patients with acute myocardial infarction within 12 hours after the onset of the chest pain, 30 patients with unstable pectoris and 30 patients with stable angina pectoris were included. Al patients were successfuly treated by percutaneous coronary intervention. Meanwhile, 30 patients only undergoing coronary angiography but not stenting treatment were selected, and another 30 patients without any treatment served as normal control group. RESULTS AND CONCLUSION:According to enzyme-linked immunosorbent method, the visfatin levels of acute myocardial infarction, unstable angina, stable angina and coronary angiography groups continue to rise at pre-operation, 30 minutes, 6 hours, 12 hours, 24 hours after operation, al of which were higher than that in the normal control group (P < 0.05). The results confirmed that within 24 hours after coronary stent implantation the visfatin levels continue to rise.
8.The clinical significance of typical reflux symptoms in diagnosing gastroesophageal reflux disease
Lin LIN ; Liya ZHOU ; Ye WANG ; Shifang LU ; Yaopeng ZHANG ; Shigang DING ; Sanren LIN
Chinese Journal of Internal Medicine 2014;53(7):517-520
Objective To explore the clinical significance of typical reflux symptoms in the diagnosis of gastroesophageal reflux disease (GERD).Methods Consecutive patients older than 16 years,who initially visited department of gastroenterology at clinic of Peking University Third Hospital from May 9,2012 to Dec 31,2012,were required to complete a self-reported GERD questionnaire.Upper endoscopy was performed in some selected patients.Results A total of 18 987 patients were enrolled with a response rate of 91.5%.The prevalence of symptom-defined GERD was 13.6% (2 579/18 987).A total of 4 357 (22.9%) patients underwent the upper endoscopy,and the diagnostic rates of reflux esophagitis,Barrett's esophagus,peptic ulcer disease,and upper gastrointestinal malignancy were 13.1% (572/4 357),1.8% (78/4 357),10.5% (456/4 357),and 1.7% (75/4 357),respectively.The incidence of reflux esophagitis was 22.7% (216/951) in patients with reflux symptoms and 10.5% (356/3 406) (P <0.001) in patients without reflux symptoms,2.7% (26/951) and 1.5 % (52/3 406),respectively (P =0.013) for Barrett's esophagus; 6.8% (65/951) and 11.5% (391/3 406),respectively (P<0.001) for peptic ulcer disease; 1.7% (16/951) and 1.7% (59/3 406),respectively (P =0.917) for upper gastrointestinal malignancy.Conclusions GERD is one of the major diseases at gastroenterology clinic.Typical reflux symptoms suggest a diagnosis of GERD.But some patients with peptic ulcer disease or upper gastrointestinal malignancy can also present typical reflux symptoms.Upper endoscopy is valuable to avoid the misdiagnosis of other disorders.
9.A clinical study of mechanical ventilation in the treatment of acute respiratory failure following abdominal surgery
Shifang DING ; Wei ZHOU ; Qian ZHAI ; Xiaomei CHEN ; Kefu WANG ; Chen LI
Chinese Journal of General Surgery 2001;0(08):-
Objective To explore the predisposing factors in the development of acute respiratory failure after abdominal surgery and the factors affecting the therapeutic effect of mechanical ventilation. Methods A (retrospective) study was undertaken for acute respiratory failure after abdominal surgery in 91 patients. The (underline) diseases, introducing causes and efficacy of mechanical ventilation were retrospectively analysed. (Results) Postoperative pneumonia was the cause of acute respiratory failure in 53 cases and ARDS caused by severe abdominal infection and severe acute pancreatitis in 38 cases. Of the 91 cases, complicated with COPD in 38 cases, severe malnutrion 32 cases, and hypokalemia 14 cases. Respiratory failure occurred at(4.08?2.45)days after operation. The duration of mechanical ventilation was(21.66?21.42)days; 33 cases died, and 58 cases were successfully recovered with mechanical ventilation.Conclusions The (management) of acute respiratory failure after abdominal asurgery should be rational use of mechanical (ventilation), adjustment of weaning strategy and avoidance of dependance on mechanical ventilation. Timely treatment of the primary disease, effective control of abdominal infection and aggressive symptomatic and (supportive) treatment are factors that affect the success or failure of mechanical ventilation.
10.Analysis of central venous catheter related sepsis
Shifang DING ; Wei ZHOU ; Enhua SUN ; Xiaojun SUI ; Xiaomei CHEN ; Kefu WANG ; Shen LI
Parenteral & Enteral Nutrition 1997;0(04):-
Objectives: To investigate the distribution of common pathogens and their antibiotic resistance from patiens with catheter related sepsis (CRS).Methods: Catheter bacteria cultrure and antibiotic sensitivity test were performed from 69 patiens with CRS.Results: The common pathogens in CRS were fungi (41.1%),Gram-positive cocci (35.6%)and Gram-negtive bacilli (23.3%). Non-C. albicans species were major pathogen (19/30 stranins).The most strains were staphylococcus epidermidis in Gram-positive cocci and the most of them were Methicillin resistant.No vancomycin resistant strains were found. The Gram negative bacilli were often resistant to third generation cephalosporens.Conclusions: The dorminant pathogens of CRS are fungi and gram positive cocci and we should pay more attention to pathogens of resistence to antibiotics. In order to control CRS, CVC must be used reasonably and shorten the duration of retention.