OBJECTIVETo study the inhibitory effects of insulin on nuclear factor-kappa B (NF-kappaB) nuclear translocation of vascular endothelial cells induced by burn serum and its correlative mechanism.

METHODSHuman umbilical vein endothelial cells (HUVECs) were cultured in vitro and divided into 5 groups: blank control group (BC, ordinary culture without any stimulation), normal serum control group (NS, cultured with nutrient solution containing 20% healthy human serum), burn serum stimulation group (BS, cultured with nutrient solution containing 20% burn human serum), burn serum+insulin treatment group (BI, cultured with nutrient solution containing 20% burn human serum and 1x10(-7) mol/L insulin), inhibitor pretreatment group [IP, pretreated with 50 micromol/L protein kinase B (Akt) specific inhibitor LY-294002, then cultured with the same medium as used in BI group 30 minutes later] according to the random number table. Six hours later, the injury and apoptosis of HUVECs was respectively observed by the scanning electron microscope and determined by the flow cytometry. Meanwhile, the phosphorylation of inhibitor kappa B-alpha (p-IkappaB-alpha) and Akt (p-Akt) in cytoplasm, and the content of NF-kappaB-p65 in nucleus were determined with Western blot.

RESULTS(1) Compared with those in BC group, HUVECs in BS group shrank obviously with irregular nuclear structure, and intercellular links jagged or vanished. Slight change was observed in HUVECs structure in NS and BI groups, with the cell ductility and nuclear structure much better than those in BS group. (2) The apoptosis rates of HUVECs in BS group [(28.5+/-2.3)%], BI group [(22.3+/-1.8)%], and IP group [(29.7+/-2.4)%] were all obviously higher than that in BC group [(15.7+/-2.2)%, F=14.288, P<0.05 or P<0.01]. There was no significant statistical difference between NS group [(17.0+/-2.5)%] and BC group in apoptosis rate (F=14.288, P>0.05). The apoptosis rate of HUVECs in BI group was obviously lower than that in BS group (F=14.288, P<0.05). (3) Compared with those in BC group, the protein expressions of p-IkappaB-alpha in cytoplasm and NF-kappaB-p65 in nucleus were up-regulated, and the protein expression of p-Akt in cytoplasm was down-regulated in BS and IP groups. The expression levels of the three proteins in NS and BI groups were close to those in BC group.

CONCLUSIONSInsulin could inhibit the IkappaB phosphorylation, and then restrict NF-kappaB nuclear translocation and improve the vascular endothelial cells function accordingly through regulating phosphatidylinositol 3 kinase/Akt pathway.

Apoptosis ; Burns ; blood ; Cells, Cultured ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; cytology ; metabolism ; Humans ; I-kappa B Proteins ; metabolism ; Insulin ; pharmacology ; NF-kappa B ; metabolism ; Phosphorylation ; Serum ; metabolism ; Umbilical Veins ; cytology

OBJECTIVETo study the effects of different perioperative treatments on the number and proportion of gut flora in SD rats.

METHODSForty-eight SD rats were randomized into 8 groups including the control group, antibiotics group, bowel preparation group, fasting group, antibiotic-bowel preparation group, antibiotics-bowel preparation-fasting group, bowel preparation-surgery-antibiotics-early postoperative feeding group (early feeding group), and bowel preparation-surgery-antibiotics-postoperative fasting group. The rats were sacrificed and stool specimens were collected from the cecum. Stools were diluted and transferred to selective medium. Bacteria counts were calculated after 48 hours of culture under constant temperature. The changes in gut flora between the different groups were compared in terms of E.coli, Bacteroides, Bifidobacterium, and Enterococcus.

RESULTSCompared with the control group, the total bacteria, Bacteroid, Enterococcus, Bifidobacterium were all significantly decreased(P<0.05), while the E.coli count and the bacillus/coccus ratio were significantly elevated(P<0.05). In the bowel preparation group, the total bacteria count, Bacteroid, Enterococcus, Bifidobacterium were all significantly decreased(P<0.05), while the E.coli count remained stable(P>0.05) and the bacillus/coccus ratio was significantly elevated(P<0.05). In the fasting group, the total bacteria count, Bacteroid, Enterococcus, Bifidobacterium were all significantly decreased(P<0.05), while the E.coli count remained stable(P>0.05) and the bacillus/coccus ratio was significantly elevated(P<0.05). Early postoperative feeding increased E.coli, Enterococcus, and total bacteria count(P<0.05), and lowered bacillus/coccus ratio(P<0.05) as compared to the fasting group.

CONCLUSIONSAntibiotics, bowel preparation, and fasting have influence on the gut flora of SD rats in count and bacillus/coccus ratio, leading to dysbiosis. Early postoperative feeding may improve dysbiosis.

Animals ; Feces ; microbiology ; Male ; Microbiota ; Perioperative Care ; methods ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effects of the different treatment on gut flora in patients with rectal cancer in the perioperative period.

METHODSA total of 64 patients with rectal cancer were prospectively enrolled from July 2010 to June 2011 at the Qingdao University Medical College Affiliated Hospital, and randomized into 8 groups receiving different treatments in perioperative period. Factorial design was used to study three factors including preoperative bowel preparation, antibiotics use, and postoperative fasting. Patients were randomized into 8 groups with 8 patients in each group using the random digit table. Preoperative and postoperative stool specimens were collected and diluted, which were transferred to selective medium. Bacteria counts were calculated after 48 hours of culture under constant temperature. The changes in gut flora between the different groups were compared.

RESULTSCompared to the preoperative parameters, total bacteria, Bifidobacterium, Peptostreptococcus, Lactobacillus, Bacteroides, Enterococcus decreased significantly(P<0.05), while the E.coli count increased significantly. The bacillus/coccus ratio was significantly imbalanced. Preoperative bowel preparation, oral antibiotics, and postoperative fasting were all predominant factors associated with gut flora(all P<0.05). Compared with the antibiotic restriction group, Bacteroides, Enterococcus, Lactobacillus, Peptostreptococcus, and total bacteria count were reduced significantly, and the bacillus/coccus ratio increased in the non-antibiotics restriction group(P<0.05). In the bowel preparation group, Bacteroides, Peptostreptococcus, total bacteria count, and the bacillus/coccus ratio decreased(P<0.05). In the postoperative fasting group, Bacteroides, Enterococcus, total bacteria, and bacillus/coccus ratio decreased(P<0.05).

CONCLUSIONSAntibiotics, bowel preparation, and postoperative fasting can affect the number and ratio of gut flora in patients with rectal cancer in the perioperative period, leading to dysbacteriosis.

Feces ; microbiology ; Humans ; Microbiota ; Perioperative Care ; adverse effects ; Prospective Studies ; Rectal Neoplasms ; microbiology ; surgery ; Risk Factors

OBJECTIVETo compare the effect of different antegrade pulmonary blood flow on the further development of pulmonary artery after Glenn procedure in cyanotic congenital heart defects (CHD) patients.

METHODSBetween October 2000 and December 2006, 132 CHD patients with decreased pulmonary artery blood flow underwent bidirectional Glenn shunt, among them 18 patients received intraoperative lung biopsy. Patients were divided into two groups according to their different sources of antegrade pulmonary blood flow: antegrade arterial blood flow group (n = 33) and antegrade venous blood flow group (n = 99). The percutaneous oxygen saturation (SpO2), hemoglobin (Hb) concentration, and hemotocrit (Hct) value were examined and recorded before and after operation. The diameters of left pulmonary artery (LPA) and right pulmonary artery (RPA) were measured with two-dimensional echocardiography and the results were used to calculate the pulmonary artery index (PAI). The method of half-quantitative morphometric technique and an image analyzer were used to measure the following indicators of pulmonary microvessels: the percentage of media thickness (MT%), the percentage of media section area (MS%), vascular numbers of per square centimeter (VPSC), and mean alveolar number (MAN).

RESULTSBefore the operation, obvious cyanosis was found in both groups, while SpO2, Hct, and Hb were not significantly different (P > 0.05). LPA, RPA, and PAI were not significantly different between two groups (P > 0.05). The MT% and MS% in antegrade venous blood flow group were significantly less than those in antegrade arterial blood flow group (P < 0.05), but VPSC and MAN were not significantly different (P > 0.05). After Glenn procedure, hypoxia and cyanosis were remarkably improved in both two groups. There was a significantly negative correlation between SpO2 and Hct (r = -0.49, P < 0.01) or Hb (r = -0.196, P < 0.01 ). The PAI increased by 22% in antegrade arterial blood flow group and 44% in antegrade venous blood flow group (P < 0.05). The diameters of LPA and RPA in antegrade venous blood flow group were significantly larger than those in antegrade arterial blood flow group (P < 0.05) and the growth of RPA in antegrade arterial blood flow group was not significant.

CONCLUSIONA better pulmonary artery growth occurs in the patients of pulmonary stenosis after Glenn shunt than in those of pulmonary atresia, and it contributes to an earlier completion of Fontan procedure.

Blood Flow Velocity ; Cardiac Surgical Procedures ; Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; physiopathology ; surgery ; Humans ; Infant ; Male ; Pulmonary Artery ; growth & development ; physiopathology ; surgery ; Pulmonary Veins ; physiopathology ; Treatment Outcome

OBJECTIVETo investigate the relationship between pulmonary pathological features and clinical physiology of congenital heart defects (CHD) with decreased pulmonary artery blood flow.

METHODSBetween July 2001 and May 2006, 18 patients with CHD with decreased pulmonary artery blood flow undergoing palliative or definitive repair and having lung biopsy intraoperatively were enrolled in this study. The patients' age was 0.4 - 8.0 years, and body weight was 6.0 - 20.0 kg. The method of semi-quantitative morphometric technique and an image analyzer were applied to measure the following indices of pulmonary microvessels: the percentage of media thickness (MT%), the percentage of media section area (MS%) and numbers of microvessels per square centimeter (VPSC). The diameters of left pulmonary artery (LPA) and right pulmonary artery (RPA) were measured with two-dimensional echocardiography. The percutaneous oxygen saturation (SpO(2)), hemoglobin concentration (HB) and hematocrit value (HCT) were examined and recorded preoperatively.

RESULTSThere was a significant negative correlation between SpO(2) and HCT or Hb (R(2) = 0.4914, P = 0.001 and R(2) = 0.5505, P < 0.001), the variation trend of these three variables was linked. There was a negative correlation between SpO(2) and the body weight (R(2) = 0.2208, P = 0.049), which is in accordance with clinical features of aggravated process of cyanosis and hypoxia. The morphological observation of lung biopsy specimens indicated that most of peripheral pulmonary arteries were distended, irregular and their walls were uneven, and "lake" type of pulmonary AV malformations were observed. There was a positive correlation between VPSC and the body weight or BSA (R(2) = 0.5472, P < 0.001 and R(2) = 0.5233, P = 0.001). There was a significant correlation between VPSC and LPA or RPA (R(2) = 0.4312, P = 0.003 and R(2) = 0.2463, P = 0.036). It was shown that the diameter of central pulmonary arteries could be a reflection of peripheral pulmonary artery growth. The diameter of LPA also correlated with the diameter of RPA (R(2) = 0.286, P = 0.022).

CONCLUSIONSFor patients with congenital heart defects with decreased pulmonary blood flow, the pulmonary pathological changes are the bases of their clinical physiologic features. It is suggested that they should be treated in their earlier stage of life.

Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; pathology ; physiopathology ; Humans ; Infant ; Lung ; blood supply ; Male ; Pulmonary Artery ; abnormalities ; pathology ; physiopathology

Objective To design an outpatient appointment system based on ASP.NET which gains advantages over the existing system in database stability,functional module integrity and interface friendliness.Methods The ASP.NET-based system was developed with Microsoft Visual Studio 2010 as the platform,Microsoft SQL Server 2005 to create data and kinds of components to establish database and application interface.Results The system enhanced hospital registration and patient satisfaction.Conclusion The system is worthy promoting for outpatient appointment in the hospital.

OBJECTIVE: To study the characteristics and biomechanical mechanism of riding injuries involving bicycles collided by motor vehicles. METHODS: The real traffic accident cases of bicycles collided by motor vehicles, including the information of scenes, bicycles, motor vehicles, rider wounds and traffic directions, were collected. Retrospective method was used to study these riding injuries. In addition, typical cases were selected to simulate traffic accident courses with computer simulation software, and the dynamic data like acceleration, force, moment were cxtracted to compare with those in the real cases. RESULTS: There were no difference of occurring frequency between cases with or without riding injuries, as well as between one-side-collision and front- or back-collision. The riding injuries seemed less in accidents involving large-scale vehicles. The frequency of riding injuries increased with vehicle speed. The wound location was low on collision side and high on opposite. CONCLUSION Analysis of riding injury characteristic in traffic accidents and their biomechanical mechanism would be helpful for estimation of traffic manner.
Accidents, Traffic ; Area Under Curve ; Bicycling/injuries* ; Biomechanical Phenomena ; Computer Simulation ; Humans ; Leg Injuries/pathology* ; Models, Theoretical ; Motor Vehicles ; Perineum/injuries* ; Retrospective Studies ; Wounds and Injuries/pathology*

OBJECTIVE: Using computer simulation to analyze the effects of speed, type of automobile and impacted position on crash-course and injuries of pedestrians in automobile vs. pedestrian accidents. METHODS: Automobiles (bus, minibus, car and truck) and pedestrian models were constructed with multi-body dynamics computing method. The crashes were simulated at different impact speeds (20, 30, 40, 50 and 60 km/h) and different positions (front, lateral and rear of pedestrians). Crash-courses and their biomechanical responses were studied. RESULTS: If the type of automobile and impact position were the same, the crash-courses were similar (impact speed < or = 60 km/h). There were some characteristics in the head acceleration, upper neck axial force and leg axial force. CONCLUSION Multi-body dynamics computer simulation of crash can be applied to analyze crash-course and injuries (head, neck and leg) of pedestrians.
Accidents, Traffic ; Automobiles ; Biomechanical Phenomena ; Computer Simulation ; Humans ; Models, Biological ; Walking

The molecular pathogenesis of leukemia is poorly understood. Earlier studies have shown both Wilms' tumor 1 suppressor gene (WT1) and CML28 abnormally expressed in malignant diseases of the hematopoietic system and WT1 played an important role in leukemogenesis. However, the relationship between molecular CML28 and WT1 has not been reported. Here we described the use of small interfering RNA (siRNA) against WT1 and CML28 in leukemic cell line K562 to examine the interaction between CML28 and WT1. WT1 and CML28 gene expression in transfected K562 cells was detected by using RQ-PCR and Western blotting. K562 cells transfected with WT1-siRNA could greatly decrease both mRNA and protein expression levels of WT1 and CML28. In contrast, CML28-siRNA did not exert effect on WT1. Further, subcellular co-localization assay showed that the two proteins could co-localize in the cytoplasm of K562 cells, but WT1/CML28 complexes were not detected by using immunoprecipitation. It was suggested that there exists the relationship between CML28 and WT1. CML28 may be a downstream target molecule of WT1 and regulated by WT1, which will provide important clues for further study on the role of CML28 and WT1 in leukemic cells.
Antigens, Neoplasm ; metabolism ; Cell Line, Tumor ; Exosome Multienzyme Ribonuclease Complex ; metabolism ; Humans ; K562 Cells ; Leukemia, Erythroblastic, Acute ; metabolism ; Neoplasm Proteins ; metabolism ; Protein Interaction Mapping ; RNA-Binding Proteins ; metabolism ; Subcellular Fractions ; metabolism ; WT1 Proteins ; metabolism

This study was aimed to investigate the expression of GST-CML28 in Escherichia Coli and to prepare its antibody. The constructed recombinant expression vectors CML28-pGEX-3X were transformed into Escherichia Coli BL21 under IPTG induction. The protein was abstracted from the transformers, and purified by a GSTrap FF column. The rabbits were immunized by the purified fusion protein to produce serum with anti-CML28 antibody. The serum was purified by chromatographic column stuffed with glutathione Sephamse 4B to get the antibody. The specific antibody against CML28 was further identified by ELISA, Western blot, immunohistochemistry and quantum dot luminescence. The results indicated that GST-CML28 fusion protein was expressed in Escherichia coli and its specific polyclonal antibody was obtained. It is concluded that the anti-CML28 polyclonal antibodies with high titer and specificity are successfully prepared. These antibodies provide an useful experimental tool to profoundly research the physiological significance and biological function of the CML28 gene.
Animals ; Antibodies ; immunology ; isolation & purification ; metabolism ; Antigens, Neoplasm ; biosynthesis ; immunology ; isolation & purification ; Cells, Cultured ; Escherichia coli ; metabolism ; Exosome Multienzyme Ribonuclease Complex ; biosynthesis ; immunology ; isolation & purification ; Genetic Vectors ; Glutathione Transferase ; biosynthesis ; isolation & purification ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; RNA-Binding Proteins ; biosynthesis ; immunology ; isolation & purification ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; immunology ; isolation & purification