Objective To explore the protective effect of L-carnitine on myocardial systolic dysfunction in sepsis and its underlying mechanism.Methods A mouse sepsis model was established by cecal ligation and puncture(CLP).Ten-week-old male SPF-grade C57BL/6 mice(body weight 20～30 g)were randomly divided into 5 groups via random number table:Sham group,Sepsis group,L-carnitine group,L-carnitine+Etomoxir(Eto)group,and Eto group.Echocardiography assessed cardiac function,ELISA measured serum creatine kinase isoenzyme MB(CK-MB)levels,and 72-hour survival rates were recorded to evaluate L-carnitine's effects on cardiac function.Cardiomyocytes were isolated,and a cell microtensiometer was used to detect cardiomyocyte contractile function and calcium transients.Myocardial tissues were collected from each group,and ELISA was used to determine the contents of triglyceride(TG),free fatty acid(FFA),and adenosine triphosphate(ATP).An in vitro sepsis model was constructed by stimulating HL-1 cardiomyocytes with lipopolysaccharide(LPS)for 12 hours,which was divided into 5 groups:control(CTRL)group,LPS group,L-carnitine group,L-carnitine+Eto group,and Eto group.ELISA was used to detect the contents of TG,FFA,and ATP as well as the activity of carnitine palmitoyltransferase 1A(CPT1A)in cardiomyocytes.A cellular energy metabolism analysis system was employed to measure fatty acid oxidation capacity,and Western blot was used to detect the protein expression of CPT1A in cardiomyocytes.BODIPY-FL-C16(green fluorescently labeled palmitic acid)was utilized to detect the distribution of fatty acids in the cytoplasm and mitochondria via immunofluorescence technology,thereby observing the ability of cells to transport fatty acids into mitochondria.Results Compared with the Sham group,cardiac function was significantly impaired in the Sepsis group,as evidenced by decreased ejection fraction and mean arterial pressure(P<0.05),along with elevated levels of the cardiac injury marker CK-MB(P<0.05).Treatment with L-carnitine significantly improved myocardial function,restored blood pressure in septic mice,and increased their survival rate from 12.50%to 81.25%(P<0.05).Compared with the Sham group,the contractile function and calcium transients of acutely isolated single cardiomyocytes were significantly reduced in the Sepsis group(P<0.05),while L-carnitine treatment remarkably restored the contractile function and calcium release capacity of septic cardiomyocytes(P<0.05).Both in vivo and in vitro experiments showed that TG and FFA levels were significantly increased(P<0.05),and ATP levels was significantly decreased(P<0.05)in the Sepsis and LPS groups—effects significantly reversed by L-carnitine treatment.Compared with the CTRL group,the basal oxidation rate and maximum oxidation capacity of fatty acids in cardiomyocytes of the LPS group were significantly reduced(P<0.05),and L-carnitine treatment notably improved these indicators.Compared with the CTRL group,the expression and activity of CPT1A in cardiomyocytes of the LPS group were significantly decreased(P<0.05),while L-carnitine treatment significantly increased the expression and activity of CPT1A(P<0.05).In LPS group cardiomyocytes,green fluorescently labeled palmitic acid primarily formed numerous granular/clumpy aggregates in the cytoplasm with minimal mitochondrial colocalization.In the L-carnitine group,the green fluorescent granules in the cytoplasm of cardiomyocytes were smaller,and colocalization with mitochondria was increased.However,the L-carnitine+Eto group exhibited similar phenomena to the LPS group.In addition,both in vivo and in vitro experiments demonstrated that treatment with the CPT1A inhibitor Eto reversed the effect of L-carnitine.Compared with the L-carnitine group,the ATP content in the L-carnitine+Eto group was significantly decreased(P<0.05),while the FFA content was significantly increased(P<0.05).Conclusion L-carnitine facilitates fatty acid entry into mitochondria for β-oxidation via a CPT1A-dependent mechanism,thereby ameliorating fatty acid oxidation dysfunction in septic cardiomyocytes and improving myocardial contractile function.

OBJECTIVEInflammation plays a critical role in secondary brain damage after intracerebral hemorrhage (ICH). However, the mechanisms of inflammatory injury following ICH are still unclear, particularly the involvement of NLRP3 inflammasome, which are crucial to sterile inflammatory responses. In this study, we aim to test the hypothesis that NLRP3 signaling pathway takes a vital position in ICH-induced secondary inflammatory damage and detect the role of N-methyl-D-aspartic acid receptor 1 (NMDAR1) in this progress.

METHODSICH was induced in mice by microinjection of hemin into the striatum. The protein levels of NMDAR1, NMDAR1 phosphorylation, NLRP3 and IL-1b were measured by Western blot. The binding of NMDAR1 to NLRP3 was detected by immunoprecipitation.

RESULTSThe expression of NMDAR1, NMDAR1 phosphorylation, NLRP3 and IL-1b were rapidly increased after ICH. Hemin treatment enhanced NMDAR1 expression and NMDAR1 phosphorylation, as well in cultured microglial cells treated by hemin. Hemin up regulated NLRP3 and IL-1b level, which was reversed by MK801 (NMDAR antagonist) in vitro. Hemin also promoted the binding of NMDAR1 to NLRP3.

CONCLUSIONOur findings suggest that NMDAR1 plays a pivotal role in hemin-induced NLRP3-mediated inflammatory damage through synergistic activation.

Animals ; Cells, Cultured ; Cerebral Hemorrhage ; complications ; Hemin ; pharmacology ; Inflammation ; etiology ; Mice ; Mice, Inbred C57BL ; NLR Family, Pyrin Domain-Containing 3 Protein ; physiology ; Phosphorylation ; Receptors, N-Methyl-D-Aspartate ; physiology ; Signal Transduction

Objective To explore the influence of drug dosage, solvent and other main influencing factors on the successful establishment of alloxan-induced hyperglycemia mouse model and the effect on the stability of this model. Methods 160 6-8-week-old Kunming mice ofSPF grade, (male:female=1:1) were used in this study.The influences of different dosages of alloxan and solvent combinations on the successful establishment rate of the model, survival rate, body weight, fasting blood glucose, blood glucose area under curve, serum insulin level and their stabilities were dynamically observed for six weeks.Results By single intraperitoneal injection of 160 mg/kg bw alloxan ( pH 4.5 citrate sodium as solvent) , we were able to obtain a stable experimental hyperglycemic mouse model with higher levels of successful establishment rate (70%), survival rate (75%), fasting blood glucose (15-20 mmol/L), glucose area under the curve (55-65 mmol/L) and a lower but not loss of serum insulin levels (21 mIU/L).Conclusions In the present study we have carefully considered the influence of main factors such as drug dosages, solvent, etc., on the alloxan-induced experimental hyperglycemic mouse model, and successfully established this model after 6-week period observation of its stability.This model may provide a useful tool in the research of experimental diabetes and hypoglycemic functional studies.

OBJECTIVETo distinguish non involuting congenital hemangioma (NICH) and infantile hemangioma (IH) by comparing the pathological structure and marker antigen expression.

METHODSFrom Jan. 2005 to Aug. 2010, 39 paraffin-embedded samples, including 13 cases of NICH, 13 cases of proliferating IH and 13 cases of involuting IH, were collected from operation. Hematoxylin-eosin staining was used to observe the pathological structure. Immunohistochemical analysis was also performed to investigate the expression of Glut-1.

RESULTSThe lobules of capillaries were well-defined in NICH. The lobules were surrounded by abundant fibrous tissue. The capillaries were often large and integrity in NICH. There were few mitosis and apoptosis in endothelial cells and stromal cells in NICH. While in IH, the pathologic findings were totally different. Immunochemistry revealed that the Glut-1 was expressed in endothelial cells of IH, but not in NICH.

CONCLUSIONSNICH has a steady histologic structure and low proliferation, while the endothelial cells in proliferative IH has a high proliferation. Glut-1 can be used as the reliable marker antigen for differential diagnosis of NICH and proliferative infantile hemangiomas.

Child ; Child, Preschool ; Diagnosis, Differential ; Female ; Glucose Transporter Type 1 ; metabolism ; Hemangioma ; congenital ; diagnosis ; pathology ; Humans ; Infant ; Male

OBJECTIVETo establish a three-dimentional liver function evaluation system using 99mTc-diethyl iminodiacetic acid (99mTc-EHIDA) scintigraphy based on single photon emission computed tomography (SPECT).

METHODSTotally 16 patients with liver lesions were divided into cirrhosis group and non-cirrhosis group. SPECT was performed 2 days before operation and 5 days after operation. Serum liver functions were examined on the same day of scintigraphy. SPECT images of areas of interest of heart and liver were aquired. Time of the peak of EHIDA density in liver (Tpeak), five-minutes heart liver index (HLI5), blood clearance index (HH15), receptor index (LHL15), and the predictive values were calculated.

RESULTSTpeak was not significantly different between two groups, while HLI5, HH15, and LHL15 were significantly different (P = 0.033, P = 0.001, and P = 0.005). HLI, and LHL15 were significantly correlated with preoperative total protein and prealbumin levels (P = 0.003, P = 0.015, P = 0.022, P = 0.038) and post-operative prealbumin (P = 0.037, P = 0.042). The predictive values of HLI5 and LHL15 correlated well with postoperative HLI5 and LHL15 (r = 0.675, P = 0.016; r = 0.629, P = 0.028).

CONCLUSIONThe three-dimentional liver function evaluation system using 99mTc-EHIDA based on liver SPECT may facilitate the further studies of risks of liver surgery.

Adult ; Aged ; Animals ; Female ; Humans ; Liver Diseases ; diagnosis ; diagnostic imaging ; physiopathology ; Liver Function Tests ; Male ; Middle Aged ; Postoperative Period ; Preoperative Period ; Radiopharmaceuticals ; administration & dosage ; Technetium Tc 99m Diethyl-iminodiacetic Acid ; administration & dosage ; Tomography, Emission-Computed, Single-Photon

OBJECTIVETo investigate the expression and role of glucose transporter-1 (Glut1) in infantile hemangioma.

METHODSFifty-two samples from infantile hemangioma, 25 in cavernous venous malformation, 9 in arteriovenous malformation, 2 in capillary malformation and 5 in normal skin samples were involved in this study. The EnVision immunohistochemical stain was used to investigate the expression of Glut1 protein in these samples.

RESULTSIn the early proliferating stage, a number of endothelial cells expressed Glut1. In the middle proliferating stage, most of vascular endothelial cells and scattered endothelial cells expressed Glut1. In the late proliferating stage, the expression of Glut1 decreased quickly. In the involuting stage, all hemangioma samples didn't express Glut1. All of the samples from the cavernous venous malformations, arteriovenous malformations, capillary malformations and normal skin had no expression of Glut1.

CONCLUSIONSGlut1 may be one of the phenotypes of infantile hemangioma endothelial cells in their development, rather than the inherent character. The expression of Glut1 changes according to the metabolic need of infantile hemangioma cells.

Blood Vessels ; Child ; Child, Preschool ; Endothelium, Vascular ; metabolism ; Glucose Transporter Type 1 ; metabolism ; Hemangioma ; metabolism ; Humans ; Infant ; Phenotype ; Vascular Malformations ; metabolism ; pathology

OBJECTIVETo investigate the distribution, phenotype and development of pericytes in infantile hemangioma.

METHODSFifty-two infantile hemangioma samples were included in our study. alpha-SMA was used as the marker antigen to observe the distribution of pericytes. Transmission electron microscope and TUNEL method were used to analyze the apoptosis of pericytes.

RESULTSIn the early and middle proliferating stage, there existed many pericytes in hemangioma; Pericytes together with endothelial cells generated vasculogenesis. In the late proliferating stage, many pericytes became apoptotic. In the early involuting stage, there were only a few of pericytes around the microvessels; After that, the microvessels became obstruction progressively and pericytes disappeared finally.

CONCLUSIONSThe pericyte is one of the major constitutive cells of hemangioma. The vasculogenesis, development and disappearance of microvessels undertaken by pericytes and endothelial cells lead to the pathologic evolution of infantile hemangioma.

Child ; Child, Preschool ; Female ; Hemangioma ; pathology ; Humans ; Infant ; Male ; Microcirculation ; Neovascularization, Pathologic ; pathology ; Pericytes ; pathology

OBJECTIVETo improve sternal elevation for pectus excavatum to be more simple, less injured and less recurrent.

METHODSWe modified procedures for the sternal elevation for pectus excavatum by dispersal of the shortened fibrous bundle connection with central tendon of the diaphragm, correction of the reverse angle of sternocostales joins, transverse cuneiform anterior osteotomy of sternum and reconnection of oblique resected costal cartilage.

RESULTSSince March 1997, 8 children (4 - 10-year-old) with the pectus excavatum have been treated by this modified sternal elevation, 4 of them who suffer from quick heart pulse improved their heart rate immediately during the operation, all patients have less bleeding and good cosmetic appearance without any complications. There were satisfactory results without recurrence after 6 months to 1 year follow-up.

CONCLUSIONThe modified sternal elevation for pectus excavatum is safe, effective and reliable method.

Child ; Child, Preschool ; Female ; Follow-Up Studies ; Funnel Chest ; surgery ; Humans ; Male ; Sternum ; surgery

OBJECTIVETo introduce our experience in diagnosis and treatment of 33 patients with Tessier craniofacial clefts.

METHODS33 patients with craniofacial clefts were classified by Tessier classification. According to the type and severity of the clefts, various techniques, from simple local flap transfer to complicated osteotomy and bone grafting were used to correct the deformity in 29 patients.

RESULTSAll patients who underwent corrective operation were satisfied with the result, and there were no complications.

CONCLUSIONS(1) Tessier classification is very important for plastic surgeon to find potential craniofacial deformities related to main signs. (2) No. 7 cleft is one of most common Tessier craniofacial clefts. (3) Each Tessier cleft is unique, therefore, the treatment plans cannot be standardized. Specific corrective operation must be performed on each patient according to the type and severity of the cleft, including simple local flap transfer to complicated osteotomy and bone grafting or distraction osteogenesis.

Craniofacial Abnormalities ; classification ; diagnosis ; surgery ; Humans

OBJECTIVETo investigate the expression of Fas/FasL in infantile hemangiomas and discuss the role of Fas/FasL in the pathologic evolution of infantile hemangioma.

METHODThe EnVision immunohistochemical stain and RT-PCR technique was used to examine the expression of Fas/FasL protein and mRNA in the infantile hemangiomas.

RESULTS(1) In the early and middle proliferating stage, a number of infantile hemangioma cells expressed Fas. In the late proliferating stage, the number of positive cells increased obviously and the expression of Fas mRNA was reaching the strongest level. In the early regressing stage the Fas still existed in some cells and after that the expression decreased quickly. (2) Up to the middle proliferating stage, there were a few of FasL(+) cells foound. In the late proliferating stage, the number of FasL(+) cells increased significantly. From the early regressing stage, the number of FasL(+) cells decreased rapidly and disappeared.

CONCLUSIONThere may exist significant correlation between the expression of Fas/FasL and the development of the infantile hemangioma cells. The apoptosis of the infantile hemangioma cells mediated by Fas/ FasL may be the major reason of the spontaneous involution of infantile hemangioma.

Apoptosis ; Child ; Child, Preschool ; Fas Ligand Protein ; metabolism ; Hemangioma ; metabolism ; pathology ; Humans ; Hyperplasia ; Infant ; RNA, Messenger ; metabolism ; Signal Transduction ; fas Receptor ; metabolism