BACKGROUND: Experiments have confirmed that there are integrin receptors on bone marrow mesenchymal stem cells (BMSCs), but it is still uncertain whether angiogenin-1(Ang-1) can promote survival of BMSCs and protest against its apoptosis via integrins. OBJECTIVE: To investigate the effect of human recombinant angiopoietin-1 on adhesion, vigour, anti-apoptosis of rat BMSCs and its mechanism of signal transduction. DESIGN, TIME AND SETTING: The cytological in vitro controlled study was performed at the Opening Laboratory, Zhongshan Hospital Affiliated to Xiamen University from January to October 2008. MATERIALS: A total of 40 clean Sprague Dawley rats were supplied by Shanghai Experimental Animal Center, Chinese Academy of Sciences. Human Ang-1 (R&D) was used in this study. METHODS: Combining density gradient centrifugation and adherence separation was used to separate, cultivate and identify rat BMSCs. At the third to sixth passages, BMSCs were used for this study. Various matrix was used to coat 24-well culture plates for 1 hour. Following washing, monoplast suspension was added. Seven groups were set up. Group 1 served as control, BMSCs were coated with phosphate buffered saline containing 0.1% bovine serum albumin. BMSCs in the groups 2, 3 and 4 were incubated with vitronectin, fibronectin and Ang-1. BMSCs in the groups 5, 6 and 7 were firstly incubated in ethylenediaminetetraacetic acid, ?1 antibody and RGD antibody for 10 minutes, and then placed in a 24-well plate, and coated with Ang-1. MAIN OUTCOME MEASURES: Adhesion, trypan blue staining method, MTT test, Hoechst staining and Annexin V/PI were used to measure biological effects of Ang-1 on adherence, activity and anti-apoptosis of BMSCs. Combined with integrin antibody and signal transduction blocker, the expression of phosphoserine/threonine protein kinase B (pAkt), total Akt, Bcl-2 and ?-actin was examined by western blotting. RESULTS: Rat BMSCs were successfully isolated. The purity of BMSCs was 98.3% using flow cytometry. Cell adhesion assay shows Ang-1 promoted the adhesion of BMSCs at 1, 2 and 3 days compared with control group (P

Objective To investigate the relationship between circadian rhythm of perioperative melatonin secretion and neuropsychological status in patients undergoing coronary artery bypass grafting (CABG) .Methods Forty male ASA Ⅱ-Ⅲ patients aged 45-60yr scheduled for elective CABG under hypothermic cardiopulmonary bypass (CPB) or off-pump were enrolled in this study. They were allocated to CPB group ( n = 20) or off-pump group (n - 20). Blood samples were taken before induction of anesthesia (T1 ), 10 min after tracheal intubation (T2), 10 min after heparinization (T3) , 2h after skin incision (T4), immediately before neutralization of heparin with protamine (T5 ), at the end of surgery after skin suture (T6 ) and every 3h after operation for 24h for determination of plasma melatonin concentration using enzyme-linked immunosorbent. Degree of depression was assessed by Self-Rating Depression Scale; anxiety by the State-Trait Anxiety Inventory and cognitive function by neuropsychological tests respectively, the day before operation, 7-10 days after surgery and 3 months postoperatively. Results In the 24 hours after operation the circadian secretion pattern of melatonin was kept in 2 patients in CPB group and 6 patients in off-pump group, but disturbed in the remaining patients in both groups. Postoperative depression scores were significantly higher than the preoperative baseline values in both groups. Anxiety scores at 7-10 days after operation were significantly higher in CPB group than those in off-pump group (P

Objective To determine the changes in perioperative plasma melatonin, cortisol and neuron-specific enolase (NSE) and neuropsychological function in patients who developed CNS complications after coronary artery bypass grafting (CABG) .Methods Three patients developed CNS complications (cerebral infarction, motor aphasia, motor and sensory aphasia) after CABG performed under hypothermic cardiopulmonary bypass (CPB) were studied. Blood samples were taken before induction of anesthesia (T1) , 10 min after tracheal intubation (T2), 10 min after heparinization (T3), at 30 min of CPB (T4) , immediately after discontinuation of CPB (T5), at closure of chest (T6), immediately after returning to ICU (T7) and every 3 h in ICU for 24 h (T8-15) for determination of plasma concentrations of NSE, cortisol and melatonin using enzyme-linked immunosorbent assay and radioimmunoassay. Cognitive function was measured before and 10 to 20 days after operation using a battery of tests, while depression and anxiety were assessed by self-rating depression scale and state-trait anxiety inventory respectively. Results The plasma NSE concentration was increased after operation in all 3 patients. The circadian rhythms of melatonin and cortisol were disturbed in patient 1 and 2 during the 24h after operation but were retained in patient 3. In patient 1 and 3 the performance of the trailing making test, the digit symbol subtest of WAIS-R and the stroop color word interference test were poor, meanwhile they became more depressed and less anxious. Conclusion In patients who develop CNS complication after CABG, there are cognitive decline, sentimental disorders and disrupted circadian rhythm of melatonin and cortisol secretion.

Objective To investigate the value of renal CT arteriography in the preoperation evaluation of patients with renal tumors.Methods From Jan 2007 to June 2012,372 patients with renal tumors took renal CT arteriography (with Philips Brilliance 16) to evaluate the variations of their renal arteries.We executed plain scan first,then artery phase,venous phase scan and delaying scan by turns after injections of Omnipaque.All images were delivered to imaging workstation and reconstructed to 3D views of renal arteries to supply imageological supports for surgeons.Results Tumors in solitary kidneys were detected in 21 patients,and masses in both kindneys were detected in 4 cases.We collected data of 376 kidneys finally.We found premature branching renal arteries on 46 kidneys (12.2％) and accessory renal arteries on 74 kidneys (19.7％).Double accessory renal arteries were seen on 10 kidneys (2.7％),and there were 84 accessory renal arteries in summary.7 accessory renal arteries (7.2％) came from arteria phrenica and 77 (91.7％) came from abdominal aorta (x2 =65.68,P ＜ 0.01).We found the regions where the accessory renal arteries entered kidneys:65.5％ (55/84) in upper poles,19.0％ (16/84) in central regions and 15.5％ (13/84)in inferior poles.Routine CT scans found 44 variations of renal arteries in 218 cases who underwent surgeries,and then renal CT arteriography found 63 variations in the same cases (20.2％ vs 28.9％,x2 =4.47,P ＜ 0.05).Finally,surgeons found 60 variations of renal artries in operations in the 63 cases with variations found by renal CT arteriography before surgeries.Conclusions There is limitations of routine CT scan (plain scan and with contrast medium) in detection of renal artery variations.Renal CT arteriography can supply reliable supports for establishment of surgical plans for renal tumors.

Objective To investigate the influence of erythropoietin in the metabolism of extracellular matrix after myocardial ischemia-reperfusion injury.Methods The langendroff reperfusion system was applied to investigate the protective function of EPO in ischemia-reperfusion condition in rats.The effects of EPO on extracellular matrix were observed by Western blot and signal pathway blocker.The LVEDP and infarction area were observed at the same time.Results EPO significantly improved LVEDP [(19.8±0.2) mmHg vs (35.9±0.2) mmHg,IR group vs EPO +IR group,P＜0.05]and decreased infarction area (35.26%±7.13% vs 62.70%±7.23%,IR group vs EPO+IR group,P＜0.05).The expression of MMPs was significantly decreased and the expression of collagenase Ⅰ/Ⅲ was significantly enhanced by EPO (MMP2 53.2+2.6 vs 21.2+2.5;MMP9 57.6±3.1 vs 19.2±2.6;IR group vs IR+EPO group (P＜0.05);collagen Ⅰ 43.2±2.2 vs 11.4±2.3;collagen Ⅲ 55.3±3.2 vs 18.1 vs 2.3;IR+EPO group vs IR group (P＜0.05).This function can be inhibited by Mek-Erk inhibitor.Conclusion EPO plays a role in the metabolism of extracellular matrix by Mek-Erk signal pathway,which can protect the heart tissue from ischemia-reperfusion injury.

Objective To evaluate and characterize tinnitus patients, and provide profiles for further studying tinnitus.Methods This study included 2 171 tinnitus patients seeking treatment between Janurary 2014 and December 2015 from department of head and neck surgery,Sun Yixian Memorial Hospital,Zhongshan University.Tinnitus measurements consisted of pure tone audiometry and tinnitus matching procedures.The characteristics of tinnitus were analyzed by the essential information and auditory checks of the patients.Results Average age was 44.08±15.37 years old.Tinnitus frequently occurred between 41 to 50 years old(491 cases,22.60%).Unilateral tinnitus was more than bilateral tinnitus, while the cranial tinnitus was rare.The patients with mild or moderate tinnitus had the largest population,with higher average hearing threshold at 8 000 Hz in tinnitus ear than non tinnitus ear.The highest frequency of tinnitus matching frequency was high frequency(4~8 kHz).The first three causes for tinnitus were unknown reasons,sudden deafness and middle ear disease.Conclusion Larger population was subjective tinnitus.Tinnitus was closely related with hearing loss.The causes for tinnitus were complex.Tinnitus retraining therapy is a treatment that is worth to be popularized.

Objective To investigate the influence of deep brain stimulation (DBS) at high frequency to the bilateral nucleus accumbens on morphine-induced conditioned place preference (CPP) and relapse behaviors during extinction phase in rats. Methods Twenty adult SD rats were employed in the experiment. Through stereotactic operation, outer electrode cannula was implanted into rats' bilateral nucleus accumbens. After 5 days of rest, the morphine-dependent rat model with CPP was established through intraperitoneal morphine injection (10 mg/kg). The rats, after being randomly divided into experimental group (morphine+DBS) and control group (morphine+sham DBS), were electrically stimulated using DBS circuits. Rats in the experimental group were given high frequency electrical stimulations while the control group was given sham stimulation. The CPP score of the two groups was recorded the day after stimulation until successful extinction and then the extinction time was compared between the two groups. After successful extinction the rats were given small dose of morphine to trigger relapse within 24 hours, and the CPP score was recorded and compared between the two groups.Results Compared with the control group (six days), the experimental group (26 days) had a longer extinction time. After relapse, the retention time within the drug-paired chamber of the experimental group was (357.01±192.72) s, obviously shorter than that of the control group ((704.91±181.35) s;t=2.370, P=0.034 6). Conclusion High frequency DBS to rats' bilateral nucleus accumbens can prolong extinction time but inhibit relapse behavior.

Objective To investigate the effects of bone marrow mesenchymal stem cells administration on VEGF,SDF-1 and TGF-?1 expressions and heart function in myocardial infarcts rats.Methods MSCs were harvested from SD rats,cultured and passaged in vitro,then the cells were purified by density gradient centrifugation and adhesive-screening method.Rats were randomly divided into 2 groups.In the control group(n=8) the animals were treated with saline of the same volume as placebo.Four weeks after the injection,the VEGF,SDF-1 and TGF-?1 expressions and heart function were examined.Results Four weeks after the transplantation,Immunohistochemistry showed expression of VEGF and SDF-1 increased markedly in experiment group,and expression of TGF-?1 decreased.Heart function was improved in the experiment group(P

Objective To investigate the influence of Angiopoietin-1(Ang1)gene modified mesenchymal stem cells on the expression of adhension factors from endothelial cells,to clarify the feasibility of modulating inflammation reaction by gene modification after stem cells transplantation.Methods Genetic engineering rMSCs were constructed by lentivirally-tranduced Ang1 gene,the expressions of ICAM-1 and VCAM-1 at different time points after 20 ?g/L VEGF stimulating were investigated,the supernatants of gene modified rMSC were collected and then added into HUVEC culture with different concentrations of Ang1,the expressions of adhension factors after VEGF stimulated were observed at the same time.Results Genetic engineering rMSCs were successfully constructed,the expressions of ICAM-1 and VCAM-1 mRNA were significantly increased after VEGF stimulated,the maximal value was at 8 h,the ICAM-1 and VCAM-1 mRNA expressions were decreased after incubation with Ang1 at different concentrations,Ang1 suppressed the VEGF-stimulated protein levels of ICAM-1 and VCAM-1.Conclusion The supernatants of Ang1 gene modified stem cells can suppresse VEGF-stimulated inflammation reaction of HUVEC.

The eukaryotic expression of human arresten gene and its effect on the proliferation of in vitro cultured vascular smooth cells (VSMCs) in vitro were investigated. COS-7 cells were transfected with recombinant eukaryotic expression plasmid pSecTag2-AT or control plasmid pSecTag2 mediated by liposome. Forty-eight h after transfection, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of arresten mRNA in the cells, while Western blot assay was applied to detect the expression of arresten protein in concentrated supernatant. Primary VSMCs from thoracic aorta of male Sprague-Dawley rats were cultured using the tissue explant method, and identified by immunohistochemical staining with a smooth muscle-specific anti-alpha-actin monoclonal antibody before serial subcultivation. VSMCs were then co-cultured with the concentrated supernatant and their proliferation was detected using Cell Counting Kit-8 (CCK-8) in vitro. The results showed that RT-PCR revealed that the genome of arresten-transfected cells contained a 449 bp specific fragment of arresten gene, suggesting the successful transfection. Successful protein expression in supernatants was confirmed by Western blot. CCK-8 assay showed that the proliferation of VSMCs were inhibited significantly by arresten protein as compared with control cells (F=40.154, P<0.01). It was concluded that arresten protein expressed in eukaryotic cells can inhibit proliferation of VSMCs effectively in vitro, which would provide possibility to the animal experiments.