Objective To establish a practical,stable and high purity endothelial progenitor cells culture meth- od in vitro.Methods Human umbilical cord blood mononuclear cells were isolated by Ficoll density-gradient cen- trifugation,then plated on dishes coated with human fibronectin.After 48 hours,the nonaderent cells were collect- ed and replated onto fibronectin-coated dishes.After 7 days of culture,the cells were identified with the techniques of immunohistochemistry,immunofluorescence and flow cytometer.Results The cultured cells were small and spindle or polygonal in shape.Large numbers of typical endothelial progenitor cell colony-forming units were found,vWF and Flk-1 proteins expression were identified in more than 95% of the attached cells with 98% of them showing positive Dil-ac-LDL and FITC-UEA-1.According to the results from fluorescence-activated cell sorting (FACS),7.0%?1.8% of cells were recognized as CD_(133)~+.Conclusion Differential attachment technique is a practical and stable method for obtaining highly purified endothelial progenitor cells.

Migration of adventitial fibroblasts (AF) is involved in the neointimal formation which is one of the common pathological processes in several vascular diseases. The observation of whether the migratory response of AF from hypertensive animal is different from that of controls may provide an explanation of vascular remodeling in hypertension. We examined whether there is any difference between the migratory activity of AF derived from spontaneously hypertensive rat (SHR) and that from their normotensive counterpart Wistar-Kyoto rats (WKY). In addition, the role of osteopontin (OPN) in cell migration was also examined. Primary cultures of aortic adventitial fibroblasts were derived from SHR and age-matched WKY. Migration of fibroblasts was determined with the Transwell method. The mRNA expression level of OPN was measured by a real-time quantitative PCR. When compared with WKY-derived cells, migration of adventitial fibroblasts from SHR exhibited an increased response when stimulated by 10% serum (cell number per field 35.20+/-5.26 vs 22.2+/-3.27, p<0.05). Chemotaxis induced by 10 ng/ml bFGF showed a similar difference (cell number per field 30.23+/-4.54 vs 19.20+/-4.47, p<0.05). We also found that SHR-derived fibroblasts expressed a higher level of OPN mRNA than the cells from WKY (1863.23+/-43.91 vs 326.24+/-68.29, p<0.01). To verify if OPN is associated with the enhanced migratory ability in AF from SHR, we designed the antisense oligonucleotide of OPN. The results showed that the antisense OPN oligonucleotide significantly inhibited AF migration (cell number per field 38.60+/-5.98 vs 26.61+/-3.84, p<0.05), while sense and mismatch OPN oligonucleotide had no effect on cell migration. Therefore, the migration of adventitial fibroblasts appeared to be enhanced in cultures derived from SHR. OPN might be involved in the difference observed.
Animals ; Aorta ; cytology ; Cell Movement ; drug effects ; Cells, Cultured ; Fibroblasts ; cytology ; Hypertension ; physiopathology ; Male ; Osteopontin ; Phosphoproteins ; pharmacology ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Sialoglycoproteins ; pharmacology

OBJECTIVETo investigate how to select an appropriate surgical approach in the management of fracture and dislocation of lower cervical spine combined spinal cord injury.

METHODSThe clinical data of 54 patients of lower cervical spine fracture and dislocation were retrospectively analyzed. There were 29 cases with vertebral body compressive fracture and dislocation, 7 cases with vertebral body bursting fracture and dislocation, 3 cases with unilateral facet dislocation, 15 cases with bilateral facet dislocation. All cases were associated with spinal cord injury. According to American Spinal Injury Association (ASIA) grades, 21 cases were in A grade, 5 cases in B grade, 22 cases in C grade and 6 cases in D grade. All patients had surgical reduction, decompression, stabilization and fusion, 43 cases in anterior approach and 11 cases in posterior approach.

RESULTSAll patients were followed up in 12 to 36 months, the mean follow-up time was 18 months. There were no great vessels, trachea, esophagus or spinal cord iatrogenic injury. There were no pull-out and breakage of screws or plates. Fusion was achieved in all patients at an average of 12 weeks postoperatively. There were no pseudarthrosis or bone nonunion. Of all the patients, 96.3% were acquired completely reduction and the normal intervertebral height and lordosis were maintained. Patients with complete spinal cord had no neurologic recovery, but they felt relief from upper limb pain or numb. Incomplete spinal cord lesions improved on average 1-2 Frankel grade after surgery.

CONCLUSIONSFor lower cervical spine fracture and dislocation, an ideal anatomy reduction can be obtained with either anterior or posterior approach surgery. It is important to select a suitable surgical approach according to different types of cervical fracture and dislocation.

Adolescent ; Adult ; Aged ; Cervical Vertebrae ; injuries ; surgery ; Diskectomy ; methods ; Female ; Humans ; Joint Dislocations ; complications ; surgery ; Laminectomy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Spinal Cord Injuries ; complications ; Spinal Fractures ; complications ; surgery ; Spinal Fusion ; methods ; Treatment Outcome

OBJECTIVETo systematically assess the effect of early enteral nutrition support after gastrointestinal operation on prognosis.

METHODSThe Cochrane Library, PubMed, CBM, CNKI, Wanfang, and VIP databases were retrieved via computer system for randomized controlled trails(RCTs) with early enteral nutrition support to patients undergoing gastrointestinal operation. Quality of studies was evaluated by the Cochrane Jadad rating scale. Nutrition indexes, bowel function indices, postoperative complications, health-economics indices were collected. Meta-analysis was conducted with RevMan 5.2.

RESULTSEleven relevant RCTs studies with 1087 cases were enrolled, including 541 patients in the study group(early enteral nutrition) and 546 in the control group. Meta-analysis showed that patients in the study group had significantly higher levels of plasma albumin and prealbumin than those in the control group(WMD=2.87, 95%CI:1.03-4.71; WMD=0.04, 95%CI:0.02-0.05). The time of postoperative bowel ventilation in the study group was significantly shorter than that in the control group(WMD=4.10, 95%CI:-5.38--2.82). The postoperative complication rate in the study group was significantly lower as compared to the control group(RR=0.64, 95%CI:0.44-0.93).

CONCLUSIONEarly enteral nutrition support after gastrointestinal operation is safe and effective, which can improve the nutritional status, promote bowel function return, and reduce postoperative complication rate.

Digestive System Surgical Procedures ; Enteral Nutrition ; Gastrointestinal Diseases ; surgery ; Humans ; Postoperative Complications ; Prognosis ; Randomized Controlled Trials as Topic

OBJECTIVETo compare the immunity of morbid obesity (MO) before and after laparoscopic adjustable gastric banding (LAGB).

METHODS15 cases, with a mean body mass index (BMI) of 35.8 kg/m(2), were treated by LAGB from Jun. 2003 to Oct. 2003 in our department. Patients' immune parameters were determined preoperatively and 1, 3 and 6 months postoperatively. 15 cases with a normal BMI (23.6 kg/m(2)) were set as controls.

RESULTSBefore surgery, the MO had a significant lower level of CD(4)(+), CD(4)(+)/CD(8)(+) and a higher level of serum interleukin-2 (IL-2), Interleukin-6 (IL-6) than the controls (P < 0.01). There was a significant reduction of weight and BMI 6 months postoperatively (P < 0.01). At the same time, CD(4)(+) increased and serum IL-2 decreased significantly. But CD(4)(+)/CD(8)(+)and serum IL-2, IL-6 were still abnormal compare to the controls.

CONCLUSIONSMO may combined with an abnormal immunity. But after enough weight loss induced by LAGB, it can be partly reversed.

Adolescent ; Adult ; Female ; Follow-Up Studies ; Gastroplasty ; methods ; Humans ; Laparoscopy ; Male ; Obesity, Morbid ; immunology ; surgery ; Weight Loss

OBJECTIVETo investigate the long-term efficacy of anterior approach surgery on cervical spondylotic myelopathy and factors affecting prognosis.

METHODSThe data in 116 patients suffered from cervical spondylosis from January 1992 to December 2000 were reviewed, including 80 male cases and 36 female cases, whose age ranged from 36 to 76 years (mean, 51 years). The preoperative course of disease was 2 months to 20 years (mean, 19 months). There were 65 cases (56.0%) with single segments involved, 44 cases (37.9%) with two segments, 7 cases (6.0%) with three segments. Ninety-eight cases were onset slowly, 18 cases with no remote cause and aggravating quickly. Three kinds of surgeries were performed: anterior cervical decompression and autoiliac bone interbody fusion, anterior cervical decompression and fusion with threaded fusion cage, anterior cervical decompression and autoiliac bone interbody fusion with anterior screw-plate system. Improvement in spinal cord function was assessed using the Japanese Orthopaedic Association (JOA) scoring system, the long-term efficacy and influential factors were also analyzed.

RESULTSThe mean follow-up time was 7 years and three months (5 - 12 years). The mean preoperative JOA score was 9.34 +/- 1.81. The mean postoperative JOA score was 10.35 +/- 1.85. At the final follow-up, the JOA score was 14.09 +/- 1.90 and the recovery rate was 63.2%. Among the total patients, 27 cases were excellent, 47 cases were fine, 23 cases were good, 19 cases were poor, the fineness rate was 63.8%. The long-term efficacy of anterior approach surgery has close correlations with time of course, age of onset, preoperative spinal cord function and the number of affected segments, but has no correlations with modes of fusion and internal fixation.

CONCLUSIONSThe patients will be attentively observed while having a definite diagnosis of cervical spondylotic myelopathy. The good long-term results will be obtained after early anterior cervical decompression and fusion.

Adult ; Aged ; Cervical Vertebrae ; pathology ; surgery ; Decompression, Surgical ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Spinal Fusion ; methods ; Spinal Osteophytosis ; surgery ; Time Factors ; Treatment Outcome

OBJECTIVETo investigate whether hyperthermia can enhance the killing effect of 5- fluorocytosine (5- FC) on human colorectal carcinoma cell lines SW480 transfected with carcinoembryonic antigen (CEA) tissue- specific cytosine deaminase (CD) gene in vitro,and study its mechanism.

METHODSHuman colorectal carcinoma cell lines SW480 transfected with G1CEACDNa were cultured. The proliferated colonies were treated with the combined therapy of 5-FC and hyperthermia at a temperature of 43 degrees C for 30 min. After eight days, MTT was used to calculate the cellular survival rate,to analyze the killing effect of 5-FC combined with hyperthermia on SW480 cells transfected with CD gene. Flow cytometry was performed to analyze the cellular cycle and transmission electron microscope was used to observe the morphologic changes of SW480 cells after thermochemotherapy.

RESULTSHyperthermia combined with 5-FC had an enhanced killing effect on SW480-CEACD cells than 5-FC alone (P< 0.05, t =2.403, n=9). Flow cytometry revealed that the proportion of S stage cell increased in the group treated with hyperthermia and 5- FC (P< 0.001, t =7.158, n=6). Transmission electron microscope showed apoptosis after thermo- chemotherapy.

CONCLUSIONSHyperthermia can improve the anti- tumor effect of 5- FC on human colorectal carcinoma cell lines SW480 transfected with CD gene, and the cells were blocked at S stage of cellular cycle and apoptosis was induced following thermochemotherapy.

Cell Line, Tumor ; Cytosine Deaminase ; genetics ; Flucytosine ; pharmacology ; Genes, Transgenic, Suicide ; Genetic Therapy ; methods ; Hot Temperature ; Humans

The retinoid N-4-hydroxyphenyl retinamide (4-HPR also known as fenretinide), a synthetic derivative of all trans retinoic acid (ATRA), has shown as an efficient chemopreventive, chemotherapeutic agent and a potent inducer of apoptosis in various cancer cell types in vitro, including leukemic cells. However the mechanisms by which 4-HPR has the apoptotic effects is not completely elucidated. This study was aimed to investigate the effect of 4-HPR on several leukemic cells and explore its mechanisms of effect on U937 cells. The cell growth and proliferation experiments were performed [corrected] cell apoptosis was detected by annexin V; reactive oxygen species (ROS) and mitochondrial transmembrane potential (DeltaPsim) were determined; protein [corrected] expression was detected by Western blot. The results showed that 4-HPR inhibited the proliferation of U937 cells in a dose- and time-dependent manner. 4-HPR markedly [corrected] induced apoptosis in U937 cells, triggered the generation of ROS, induced the loss of mitochondrial transmembrane potential, decreased the expression of procaspase-8 and procaspase-3. Pretreatment of L-ascorbic acid suppressed the generation of ROS, disruption of mitochondrial potential, activation of caspases and apoptosis. It is concluded that the generation of ROS followed by the disruption of mitochondrial transmembrane potential plays an important role on 4-HPR-induced apoptosis in leukemic cells, suggesting that 4-HPR may be one of mitochondrial-targeted agents with clinical potential in treating cancer.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Caspases ; metabolism ; Dose-Response Relationship, Drug ; Fenretinide ; pharmacology ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; metabolism ; pathology ; Membrane Potential, Mitochondrial ; drug effects ; Reactive Oxygen Species ; metabolism ; U937 Cells

OBJECTIVETo investigate whether the number of circulating endothelial progenitor cells (EPCs) correlate with the severity of coronary stenosis in patients with stable coronary artery disease (CAD).

METHODS80 consecutive patients who underwent coronary angiography (exclusion of acute coronary syndrome and myocardial infarction) were enrolled. Physical examination and blood tests were performed to assess the disease severity and cardiovascular risk factors. Circulating EPCs as measured by the number of CD133/KDR double positive cells were detected by FACS.

RESULTSThe number of EPCs inversely correlated with age, creatinine clearance (Ccr) and left ventricular mass index (LVMI) (P = 0.004, 0.015, 0.014 respectively). Patients with hypertension showed significant reduction in number of EPCs compared to those without hypertension (P = 0.004). Moreover, the number of EPCs in patients with coronary artery disease was significantly lower than that of those with normal coronary artery (P < 0.01). EPCs also inversely correlated with angiographic Gensini score (n = 49, r = -0.305, P = 0.039).

CONCLUSIONSIn patients with stable CAD, the numbers of circulating EPCs correlate with the severity of CAD as well as cardiovascular risk factors.

AC133 Antigen ; Antigens, CD ; analysis ; Cell Count ; Coronary Disease ; etiology ; pathology ; Endothelial Cells ; physiology ; Female ; Glycoproteins ; analysis ; Hematopoietic Stem Cells ; physiology ; Humans ; Male ; Peptides ; analysis ; Risk Factors ; Vascular Endothelial Growth Factor Receptor-2 ; analysis

BACKGROUNDTo analyze the genetic and antigenic characteristics of human H5N1 virus isolated from the mainland of China.

METHODSThe hemagglutinin (HA) gene of human H5N1 virus were sequenced and analyzed.

RESULTSThe results of HA gene sequencing showed that all the virus isolates belong to the same group because of the high similarity, but they were different from the virus isolated from Thailand and Vietnam. The sequence data also showed that the receptor specificity and the connecting peptide between HA1 and HA2 are still avian influenza origin.

CONCLUSIONThe virus isolates from mainland of China until now belong to the same group and are different from the virus isolated from Thailand and Vietnam, and there is no evidence showing the human-avian influenza reassortant and recombination.

Animals ; Chick Embryo ; China ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A Virus, H5N1 Subtype ; classification ; genetics ; isolation & purification ; Influenza, Human ; virology ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA