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The clinical data of 288 patients with gastrointestinal perforation undergoing surgical treatment from Jul 2014 to Jul 2017 were retrospectively analyzed,among whom the surgical incision infection occurred in 112 patients(38.9％).The risk factors of the incision infections were examined with logistic regression analysis.The univariate analysis showed that preoperative albumin level (≤30 g/L),body mass index (＞24.0 kg/m2),duration of abdominal pain(＞24 h),extension of incision,preoperative shock,colostomy,preoperative antibiotic use and the operation time were associated with incision infections(P＜0.05),while the gender,age,preoperative hemoglobin level,diabetes,incision length were not associated with the incision infections(P＞0.05).The multivariate logistic regression analysis showed that the body mass index(OR=1.61,P＜0.01),gastrointestinal perforation site(colon and rectum,OR=5.60,P＜0.01),extension of incision (OR=3.94,P＜0.01) and operation time(OR=1.04,P=0.02)were independent risk factors of theincision infection.The results suggest that the full preoperative preparation,intensive treatment of underlying diseases,avoiding incision extension and shortening operation time may be able to reduce the surgical incision infections for patients with the gastrointestinal perforation.
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Clinical data of 12 patients with gastric cancer, in whom the Roux and Y space hernia developed after gastrectomy with Roux-en-Y anastomosis in our hospital from June 2010 to December 2018, were retrospectively analyzed. The clinical symptoms of patients were abdominal pain, distension and ileus. The main CT findings were torsion of mesentery with whirlpool sign, intestinal obstruction and exudants around the small bowels. During the operation it was found that small bowels herniated into the Roux and Y space in all 12 patients, the necrotic small intestines were resected in 4 patients. Ten patients were recovered, and 2 died. No recurrence was observed in all 10 patients during 3 month-follow up. The postoperative Roux and Y space hernia is a internal hernia and difficult to be diagnosed. The CT scan is valuable for diagnosis of Roux and Y space hernia; the main CT signs were swirled appearance of mesentery and small bowel obstruction. Once diagnosis is made the emergency operation is necessary.
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Clinical data of 12 patients with gastric cancer,in whom the Roux and Y space hernia developed after gastrectomy with Roux-en-Y anastomosis in our hospital from June 2010 to December 2018,were retrospectively analyzed.The clinical symptoms of patients were abdominal pain,distension and ileus.The main CT findings were torsion of mesentery with whirlpool sign,intestinal obstruction and exudants around the small bowels.During the operation it was found that small bowels herniated into the Roux and Y space in all 12 patients,the necrotic small intestines were resected in 4 patients.Ten patients were recovered,and 2 died.No recurrence was observed in all 10 patients during 3 month-follow up.The postoperative Roux and Y space hernia is a internal hernia and difficult to be diagnosed.The CT scan is valuable for diagnosis of Roux and Y space hernia;the main CT signs were swirled appearance of mesentery and small bowel obstruction.Once diagnosis is made the emergency operation is necessary.
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Objective@#To evaluate whether epicardial fat volume (EFV) is related to coronary artery calcification in patients with chronic kidney disease(CKD).@*Method@#Multi-slice computed tomography was performed in 30 healthy subjects and 120 patients with CKD. Cross-sectional tomographic cardiac slices from base to apex were traced semi-automatically using a Volume Viewer of AW4.3 off-line workstation, and EFV was measured by assigning Hounsfield units ranging from -250 to -30 HU to fat.The coronary artery calcification score was assessed by CaScoring software. High density lipoprotein cholesterol(HDL-C), low density lipoprotein cholesterol(LDL-C) and collecting the body mass index (BMI), dialysis route, history of diabetes and coronary artery disease were used to analyze the relationship between EFV and other risk factors in patients with CKD.@*Results@#There were 60.8%(73/120) male (mean age 62.8 years) and 39.2%(47/120) female (mean age 66.6 years) in the patients cohort, 73.3%(88/120) patients had coronary artery disease, 55.8%(67/120) had diabetes, 21 patients were on peritoneal dialysis and 9 on hemodialysis. EFV was apparently higher in stage 4-5 D CKD group compared with the control group((140.03±54.71), (145.01±64.56)and (141.45±62.04) cm3 vs.(92.42±39.56)cm3, P=0.007, 0.015 and 0.001), was similar between CKD3 and control group, and EFV was significantly higher in peritoneal dialysis group than in hemodialysis group and in coronary artery disease group compared with no coronary artery disease group((140.67±70.31) cm3 vs.(105.22±61.49) cm3, P=0.002). EFV was obviously higher in diabetes group than no diabetes group((148.41±65.78) cm3 vs.(110.53±62.37) cm3, P=0.007). CACS was apparently increased in stage 3-5 CKD group compared with the control group(140.0 vs.4.3, P<0.001). (3)When the patients were divided into four groups according to the eGFR, EFV was positively associated with CACS(rs=0.539, P=0.004) in control group, and the association become more robust in patients with CKD5(rs=0.841, P<0.000 1). EFV was related to age(r=0.662, P=0.005), BMI(r=0.648, P=0.009)and HDL-C(r=-0.433, P=0.024), but not related to eGFR and LDL-C. EFV was related to CACS(r=0.427, R2=0.182 3, P<0.001). CACS was positively correlated to age and BMI (all P<0.05)and negatively correlated with eGFR(P<0.05).@*Conclusions@#Measurement of EFV may provide another useful noninvasive indicator of coronary artery calcification in CKD patients.
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Objective To study the effects of electromagnetic field (EMF) on the activation of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) pathways in mesenchymal stem cells (MSCs) and their interaction, and to explore the cellular signal transduction mechanism of the biological effects induced by EMF.Methods The 3rd-passage rat bone marrow MSCs were randomly divided into a control group, an EMF group, an EMF + PD98059 group and an EMF + SB202190 group.Cells in the EMF group were cultured in the electromagnetic field, those in the EMF + PD98059 and EMF + SB202190 groups cultured in the electromagnetic field after PD98059 or SB202190 was added, and those in the control group were cultured normally.Different groups of cells were exposed to electromagnetic fields (15 Hz, 1 mT, sine wave form) for different exposure duration.The activated phosphorylated and non-phosphorylated p38MAPK were measured using Western blotting analysis with their specific corresponding antibodies.The alkaline phosphatase (ALP) activity in cells in different groups was detected according to the instructions of ALP kit.MTT assay was applied to investigate the proliferation of cells.Results Electromagnetic fields could rapidly induce the activation of p38 MAPK (P ＜ 0.05) and the phosphorylation of p38 MAPK elevated after 15 min exposure to EMF.The phosphorylation of p38 MAPK was significantly lower in the EMF + SB202190 group than that of the EMF group.After 5 days of EMF exposure, the ALP activity of cells was significantly improved, and the effect could be inhibited by SB202190.The bone marrow mesenchymal stem cells proliferation increased significantly after being exposed to EMF for 3 days, and it could not be blocked by SB202190.Phosphorylation of ERK and MAPK increased significantly when the p38 MAPK pathway was blocked by SB202190 and exposed to EMF for 5 minutes, and it also increased significantly when the ERK MAPK pathway was blocked by PD98059 and received 30 minutes of EMF exposure.Conclusion EMF can quickly activate ERK and p38 MAPK pathways to induce cell proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.Moreover, in EMF there is a mutual interference between ERK and p38 MAPK pathways.
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BACKGROUND: It has been demonstrated that electromagnetic field (EMF) can adjust proliferation and differentiation of bone marrow mesenchymal stem cells, but the specific mechanism is not clear. OBJECTIVE: To investigate the effects of EMF-activated ERK1/2 pathway on proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells.METHODS: The 3rd passage of rat bone marrow mesenchymal stem cells were received EMF treatment (15 Hz, 1 mT, sine wave), 20 μmol/L PD98059 + EMF treatment, or only PD98059 treatment. Simultaneously, a normal control group was established. Western blotting was applied to detect the activation of ERK signal pathway after EMF exposure. MTT assay was used to determine the activation of proliferation of cells. And alkaline phosphatase (ALP) activity in cells was detected by an ALP kit. RESULTS AND CONCLUSION: The ERK1/2 phosphorylation, proliferation and ALP activity of rat bone marrow mesenchymal stem cells were remarkably increased after exposure to EMF (P < 0.01). PD98059 could effectively block the increasing of ERK1/2 phosphorylation and cell proliferation (P < 0.01), but elevate ALP activity in a certain level (P < 0.01). EMF stimulation can fast activate ERK1/2 signal pathway and then promote the proliferation of rat bone marrow mesenchymal stem cells, however, ERK1/2 signal pathway activation has a less effect on osteogenic differentiation of bone marrow mesenchymal stem cells.
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BACKGROUND: It has been reported that in China, human bone marrow mesenchymal stem ceils are mostly harvested from adults. Studies on bone marrow mesenchymal stem cells in children are few.OBJECTIVE: To isolate and expand bone marrow mesenchymal stem cells from children, and to analyze the biological characteristics of bone marrow mesenchymal stem cells and their potential of differentiating into osteoblasts, adipocytes and neural like cells.DESIGN: Observational comparative study.SETTING: Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Experiments were performed at the Laboratory of Department of Orthopaedics of Wuhan Tongji Hospital from March to September 2006. Bone marrow mesenchymal stem cells were collected from one boy patient and two girl patients aged 5-8 years, who received pelvis osteotomy for dysplasia of the hip joint. The experimental procedures were approved by the Hospital Ethics Committee and family members of all children patients singed the informed consent.Dexamethasone, vitamin C, β-sodium glycerophosphate, 3-1sobutyl-1-methylxanthine, insulin, indometacin and butylated hydroxyanisole were bought from Sigma Company. Dimethyl sulphoxide was purchased from Amersco Company.METHODS: Bone marrow mesenchymal stem cells were cultured from mononuclear cells isolated over a Percoll gradient.Bone marrow mesenchymal stem cells were observed under an inverted phase contrast microscope. Bone marrow mesenchymal stem cells could differentiate into osteoblasts, adipocytes and neural like cells with osteoblast inductor (β-sodium glycerophosphate, dexamethasone, vitamin C), lipoblast inductor (dexamethasone, 3-isobutyl-1-methylxanthine,bovine insulin, indometacin) and serum-free medium inductor (dimethyl sulphoxide, butylated hydroxyanisole) respectively.Osteoblast marker (alkaline phosphatase, osteocalcin mRNA, calcium node), adipocyte marker (lipid droplet, PPAR γ-2mRNA) and neural ceil-like marker (nissl body, neuron specific enolase, neurofilament protein) were respectively determined by the immunohistochemical method, polymerase chain reaction and immunocytochemical method.MAIN OUTCOME MEASURES: ①Appearance and proliferation of bone marrow mesenchymal stem ceils from children,and ②determination results of osteoblast, adipocyte and neural cell markers.RESULTS: ①Children bone marrow mesenchymal stem cells could easily adhere to the wall, appeared fusiform, had high reproductive activity and arranged vortically after fusing. ②Appearance of bone marrow mesenchymal stem cells changed after receiving inductor. Osteoblast marker, adipocyte marker and neural cell-like marker were positive after chemical staining, polumerase chain reaction and immunocyte staining.CONCLUSION: Children bone marrow mesenchymai stem cells show stable proliferation, passage and multi-direction differentiation towards osteoblasts, adipocytes and neural like cells.
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BACKGROUND: It has been proved that electromagnetic field can adjust and control proliferation and differentiation of bone marrow mesenchymal stem cells v/a cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) signal transduction system. However, there are few relevant reports about Ca2+ as the second messenger in application. OBJECTIVE: To study the effects of verapamil on the proliferation and differentiation of bone marrow rnesenchymal stem cells stimulated by electromagnetic fields and to conclude influx changes of Ca2+.DESIGN, TIME AND SETTING: Electrostimulative cytological observation in vitro, which was performed in Laboratory of Orthopedic Surgery, Tongji Hospital between April and June 2005.MATERIALS: Six 4-5-week SD rats of clean grade were selected in this study. Verapami| was provided by Sigma Company, USA, and Helmholtz coil-magnetic field producer was made in Department of Electric Machine, Navy Engineering University.METHODS: The bone marrow mesenchymal stem cells were isolated and cultured in vitro with adherence method and digested with trypsin. The fourth-passage cells were harvested, adjusted to 1 × 107 L-1 in density, and divided into A, B, C and D groups in 96-well plate with 200 μ I/well. Cells in the normal control group were not performed with any agent. On the second day of inoculation, cells in the magnetic field (EMF) group were cultured in Helmholtz-coil magnetic field (0.8 mT, 50 Hz) in 0.05% CO2 saturated humidity incubator at 37 ℃, 30 minutes for each, 12 hours for interval, six time in total. Cells in the verapamil group were cultured with 20 μ mol/L verapamil, and cells in the combination group were cultured with 20 μ mol/L verapamil and magnetic stimulation.MAIN OUTCOME MEASURES: Proliferative activity was tested with MTT method, content of alkaline phosphate differentiated to osteoblasts was measured, and cells were stained with modified Gomori Ca-Co staining. RESULTS: Proliferative activity was significantly increased in the EMF group as compared with that in the normal control group after 3-day magnetic stimulation (P < 0.01), but verapamil could inhibit promotive effect on proliferation. Content of alkaline phosphate in the normal control group was similar to that in the EMF group, while those two contents were significantly higher than those in the verapamil group and the combination group (P < 0.01); furthermore, content of alkaline phosphate in the combination group was significant higher than that in the EMF group (P < 0.01). Qualitative analysis of alkaline phosphate showed a coincident result as mentioned above.CONCLUSION: EMF of 50 Hz frequency and 0.8 mT intensity can change intracellular free calcium ion concentration of bone marrow mesenchymal stem cells, and the change play a key role in the cellular proliferation and play a partial role in the differentiation of bone marrow mesenchymal stem cells into osteoblasta.
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In order to identify the differentially expressing gene of bone marrow mesenchymal stem cells (MSCs) stimulated by electromagnetic field (EMF) with osteogenesis microarray analysis, the bone marrow MSCs of SD rats were isolated and cultured in vitro. The third-passage cells were stimulated by EMFs and total RNA was extracted, purified and then used for the synthesis of cDNA and cRNA. The cRNA of stimulated group and the control group was hybridized with the rat oligo osteogenesis microarray respectively. The hybridization signals were acquired by using X-ray film after chemiluminescent detection and the data obtained were analyzed by employing the web-based completely integrated GEArray Expression Analysis Suite. RT-PCR was used to identify the target genes: Bmp1, Bmp7, Egf and Egfr. The results showed that 19 differentially expressing genes were found between the stimulated group and the control group. There were 6 up-regulated genes and 13 down-regulated genes in the stimulated group. Semi-quantitative RT-PCR confirmed that the expressions of Bmp1, Bmp7 mRNA of the stimulated group were up-regulated (P<0.05) and those of Egf, Egfr were down-regulated (P<0.05). It was suggested that the gene expression profiles of osteogenesis of the bone marrow MSCs were changed after EMF treatment. It is concluded that the genes are involved in skeletal development, bone mineral metabolism, cell growth and differentiation, cell adhesion etc.
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Células da Medula Óssea/citologia , Diferenciação Celular , Regulação para Baixo , Campos Eletromagnéticos , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Células-Tronco Mesenquimais/citologia , Hibridização de Ácido Nucleico , Sondas de Oligonucleotídeos/química , Osteogênese/genética , RNA Complementar/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Objective To screen the differential expression genes of bone marrow MSCs stimulated by electromagnetic field(EMF)with osteogenesis microarray analysis,and to study the underlying mechanism that EMF promotes the differentiation of bone marrow MSCs. Methods The Sprague-Dawley rat bone marrow MSCs were isolated and cultured in vitro.The third-passage cells who were stimulated by EMF and served as the stimulated group,and those who were not stimulated by the EMF served as the controls.Total RNA was extracted and purified,then it was used to synthesize cDNA and cRNA.The eRNA of stimulated group and the control group was hybridized with the rat oligo osteogenesis microarray,respectively.The hybridization signals were acquired by using X-ray film after chemiluminescent detection and the obtained data were analyzed using the web-based completely integrated GEArray Expression Analysis Suite.RT-PCR was used to identify the chosen genes BMP1,VDR and EGF. Results Nineteen differential expression genes were found between the stimulated group and the control group.There were 6 upregulated and 13 downregulated genes in the stimulated group.Semi-quantitative RT-PCR confirmed that the expression levels of BMP1,VDR mRNA in the stimulated group were upregulated and EGF downregulated. Conclusion The gene expression profiles about osteogenesis of the bone marrow MSCs were changed after EMF intervention(1 5 Hz,1 mT).These genes are involved in the difierentiation of bone marrow MSCs into osteoblast.These results provide deeper insight into the mechanism that EMF exposure facilitates the in vitro differentiation of bone marrow MSCs.
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Objective To study the biological effects of sinusoidal electromagnetic fields(EMFs)on proliferation and extracellular matrix(ECM)formation by annulus fibrosus(AF)cells in rats.Methods AF cells isolated from rats were randomly divided into a control group and an experimental group.The cells in the experimental group were stimulated with an EMF,while those in the control group were held under the same culture conditions but with no EMF.Flow cytometry and MTT were performed to observe the effects on the ceU cycle and proliferation.Collagen and aggrecan expression were examined after amplification with a reverse transcriptase polymerase chain reaction(RTPCR).Sulfated glycosaminoglycan(sGAG)content wag detected by applying the Alcian blue method. Results AF cell proliferation was not significant until after 4 days of stimulation.Compared with the control group,the expression of type Ⅰ and Ⅱ collagen and Aggrecan were up-regulated,and sGAG content Was increased in the experimental group.Conclusion AF cell proliferation was enhanced by EMF.Gene expression of collagen type Ⅰ and Ⅱ and Aggrecan increased.a8 well as sGAG levels.The results suggest an approach for treating of intervertebral disc degeneration.
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Objective To investigate the effects of an electromagnetic field on the extra-cellularly regulated kinase(ERK)signalling pathway and to determine the impact of electromagnetic activation on osteogenic proliferation and differentiation in rat bone marrow mesenchymal stem cells.Methods Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro.The third-passage cells were divided into 4 groups(Control,PD98059,EMF and EMF+PD98059).Western blotting Was used to detect the activation of the ERK signal pathway after exposure to an electromagnetic field.MTT assay Was used to determine the activation of proliferation in the celb in the different groups.The cells' alkaline phosphatase activities were also detected. Results (1)The ERK signal pathway in these rat bone marrow mesenchymal stem cells was activated after exposure to a 15 Hz.1 mT,sine wave form electromagnetic field for 5 min.Activation remained high for at least 1 h.PD98059 can effectively block the activation of the ERK signal pathway.(2)Cell proliferation was promoted after exposure to the electromagnetic field,and this effect could be significantly inhibited by PD98059.(3)Alkaline phosphatase was significantly elevated in these bone marrow mesenchymal stem cells after exposure to the electromagnetic field.The activation in the EMF+PD98059 group Was slightly greater than in the EMF group.Conclusion Electromagnetic fields of 15 Hz and 1 mT can activate the ERK signal pathway and alter proliferation and osteogenic differentiation in the bone marrow mesenchymal stem cells of rats.
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Objective:To investigate the changes in the expression of COX-2 in acute spinal cord injury of rats. Methods:Sixty adult SD rats were randomly divided into two groups: Trauma group (n=48) subjected to laminectomy and bounce of spinal cord; control group (n=12) only received laminectomy. The activity of COX-2 was detected using immunohistochemistry and Western blot analysis in cytoplasm of spinal cord. Results:The expression of COX-2 increased at 2-hour after injury and reached the top level at 24-hour. The expression of COX-2 dropped at 48-hour after injury and returned to basal level at 72-hour. Conclusion: The expression of COX-2 could be regarded as an index of inflammation on acute trauma spinal injury. Inhibition of COX-2 might be an new therapeutic method for spinal cord injury.
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Objective:To synthesize analogues of 6-[4-(4-substituted acyl piperazinyl)phenyl]-4, 5-dihydro-3(2H)pyri-dazinones and 6-[4-(4-substituted acyl piperazinyl)phenyl]-5-methyl-4,5-dihydro-3(2H)pyridazinones in search for more potent and selective antithrombotic drugs. Methods:Many reactions such as Friedel-Crafts reaction,hydrolysis,cyclation, acyla-tion,substitution were used to synthesize the title compounds. Born method was applied for preliminary pharmacological test in vitro. Results : Twelve title compounds were synthesized, in which 10 compounds were firstly reported. Their structures were identified by element analysis and 1H-NMR. Conclusion:Results of preliminary pharmacological test show that all synthesized compounds are effective against platelet aggregation induced by ADP in vitro. The activity of compound (9) is the most potent. The activity of compound (6), (7), (10), (11), (12) are also better than that of CCI-17810.