OBJECTIVETo investigate the protective effect of H2S pretreatment after cerebral schemia/reperfusion injury and its mechanisms in rats.

METHODSThe rat model of global cerebral ischemia/reperfusion injury was established by bilateral common carotid arteries occlusion combined with hemorrhagic hypotension.30 rats were randomly divided into four groups(1)sham group(n=5),in which rats received sham surgery only,with their bilateral vertebral artery and bilateral common carotid artery exposed but without ischemia treatment;(2)global cerebral ischemia/reperfusion model group(IR group,n=5),in which the global cerebral ischemia was induced by 10-min occlusion of bilateral common carotid arteries combined with hypotension;(3)H2S pretreatment group(n=15),in which H2S(12,24,48 Μmol/kg)was intraperitoneally injected before operation;(4)NaCl pretreatment group(n=5),in which the rats were intraperitoneally injected with saline 30 minutes before operation.The activities of superoxide dismutase(SOD)and the levels of malondialdeehyde(MDA)in brain were measured by spectrophotometry.Brain water content was detected.The expression of heat shock protein 70(HSP70) in the hippocampus was determined by Western blotting.

RESULTSThe SOD activities were significant increased in groups pretreated with 12Μmol/kg H2S(P=0.042),24Μmol/kg H2S(P=0.002),and 48Μmol/kg H2S(P=0.000),and the SOD activity was significantly lower in the ischemia group than in the Sham group(P=0.003).The MDA activities in the 24Μmol/kg group(P=0.026)and the 48Μmol/kg group(P=0.015)groups were significantly lower than in the IR group.The brain water content was decreased in H2S pretreatment group(24Μmol/kg and 48 Μmol/kg)compared with IR group(P=0.018,P=0.008),and it was also significantly higher in the IR group than in the sham group(P=0.009).The expression of HSP70 were decreased in H2S pretreatment group(24 Μmol/kg)compared with the IR group(P=0.000),and the expression of HSP70 were significantly higher in the IR group than in HSP70 group(P=0.000).The expression of HSP70 also significantly differed between NaCl group and HSP70 group(P=0.000).

CONCLUSIONH2S has protective effects on cerebral ischemia and reperfusion,which may be achieved by improving SOD activity,removing oxygen free radicals,inhibiting lipid peroxidation,and down-regulating the expression of HSP70 in the hippocampus.

Animals ; Brain Ischemia ; metabolism ; Free Radicals ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Hippocampus ; metabolism ; Hydrogen Sulfide ; administration & dosage ; therapeutic use ; Male ; Random Allocation ; Rats ; Reperfusion Injury ; metabolism ; prevention & control ; Superoxide Dismutase ; metabolism

Adult ; Dendritic Cell Sarcoma, Interdigitating ; complications ; surgery ; Humans ; Male ; Paraneoplastic Syndromes ; etiology ; Pemphigus ; etiology

OBJECTIVETo investigate the correlation between trauma and pulmonary thromboembolism.

METHODSComminuted fractures and extensive soft-tissue contusion at both hind limbs were made by a falling weight from a height in 16 rabbits. Lung perfusion scanning was performed to obtain the radioactivity counts before trauma, at 1 h, 48 h and 96 h after trauma. All the data were divided into 4 groups based on the above 4 time points. The rabbits were sacrificed when positive findings on the pulmonary perfusion scanning appeared. Their lungs were harvested to be paraffin-embedded and stained with hematoxylin-erosin method for histological examination of thromboembolism. The randomized block design ANOVA and the method of least significant difference (LSD) were used for statistical analysis of the radioactivity counts.

RESULTSThe histological findings showed that pulmonary embolism developed in 6 of the 16 rabbits (37.5%). Five of the 6 pulmonary embolism rabbits presented neither clinical symptoms nor positive pulmonary embolism manifestations in the lung perfusion scanning. A significant difference was found in lung perfusion radioactivity between the pre-traumatic, post-traumatic 1h groups and post-traumatic 48 h and 96 h groups(P less than 0.05).

CONCLUSIONSFractures of the hind limbs accompanied with extensive soft-tissue contusion may cause pulmonary micro-embolism that is not sensitive to lung perfusion scanning and tends to have no clinical symptoms. Pulmonary embolism development may take more than two days after trauma.

Animals ; Female ; Fractures, Bone ; complications ; Male ; Pulmonary Embolism ; etiology ; Rabbits ; Wounds and Injuries ; complications

Acute Disease ; Humans ; Insecticides ; poisoning ; Male ; Occupational Exposure ; adverse effects ; Poisoning ; etiology ; prevention & control

Objective To investigate a method for the purification of the N?terminal peptide fragment(NT)of the myocardial calcium channel Cav1.2,and characterize its interaction with calmodulin(CaM). Methods EscherichiacoliBL?21 cells were transformed with plasmid pGEX?6p?3/NT harboring the NT?GST fusion gene. The cells harboring pGEX?6p?3/NT were cultured and protein expression was induced with isopropyl?β?D?thiogalactoside(IPTG). Then,the GST?NT fusion protein was purified by using glutathione Sepharose 4B(GS?4B)beads. GST was cleaved off with the PreScission protease,and SDS?PAGE was performed to detect the purity and relative molecular weight of the purified peptide. Further, GST pull?down assay was performed to characterize the interaction of the NT peptide with CaM. Results SDS?PAGE analysis showed that the NT peptide was successfully purified,with high purity. Results of the GST pull?down assay showed that the NT peptide could interact with CaM. Conclusion This study establishes a method for the purification of the NT peptide and lays the foundation for further research on the interaction partners and biological functions of NT.

Objective To establish the quality standard for Sanyuan Rupixiao Gel Paste. Methods Sparganii Rhizoma, Gleditsiae Sinensis Fructus, Cyperi Rhizoma and Impatientis Semen were identified by TLC method. The content of tetrahydropalmatine was determined by HPLC. Waters symmetry column was used with the mobile phase of acetonitrile-0.1% phosphatic acid in a gradient manner (pH was adjusted to 6.4 by triethylamine) (55:45) at the detection wavelength of 280 nm. The flow rate was 1.0 mL/min at the column temperature of 30 ℃. Results The spots in TLC were clear without any interference;tetrahydropalmatine showed a good linear relation in the range of 0.092–1.84 μg;the average recovery was 100.15%with RSD of 1.58%(n=6). Conclusion The method is simple and accurate with high reproducibility, which can be used for the quality control of Sanyuan Rupixiao Gel Paste.

Allogeneic hematopoietic stem cell transplantation is the only curative therapy for severe beta-thalassemia. This time, the experience of utilizing HLA 2-loci mismatched sibling cord blood transplantation (CBT) in a child with severe beta-thalassemia was firstly reported in our country. A 3-year-male patient had been diagnosed with severe beta-thalassemia at 6 months of age (HbF 86.6%, HbA1 1.7%, HbA2 1.7%, beta globin gene mutation CD17, A-->T/IVS-II-654, C-->T). The patient's HLA typing was A 24,11, B 58,35 and DRB1 03,15. During a subsequent maternal pregnancy. The prenatal diagnosis for thalassemia and prenatal HLA typing analysis were performed on 18 weeks of pregnancy. The results indicated that the male fetus was a heterozygote (beta globin gene mutation N/CD17, A-->T), HLA typing was A 24,11, B 58,51 and DRB1 03,12. 120 ml cord blood was collected at time of delivery, the total numbers of nucleated cells, CFU-GM and CD34(+) cells were 1.830 x 10(9), 16.653 x 10(5) and 3.11 x 10(6), respectively. A new conditioning regimen including: hypertransfusion, continuous i.v. desferrioxamine, busulfan, cyclophosphamide, antithymocyte globulin plus hydroxyurea and fludarabine. GVHD prophylaxis comprised cyclosporin A and mycophenolate mofetil. The viability of cord blood at the time infusion was 92%, The total numbers of nucleated cells, CFU-GM and CD34(+) cells in the transfused cord blood were 12.06 x 10(7)/kg, 1.098 x 10(5)/kg, and 2.04 x 10(6)/kg, respectively. Results showed that the patient's clinical course after cord blood transplantation was unremarkable. Acute GVHD grade I developed on day 15, methylprednisolone 2 mg/kg was given to cure. Neutrophil engraftment (ANC > 0.5 x 10(9)/L) on day 17, platelet engraftment (> 50 x 10(9)/L) on day 50. The patients became independent from red blood cell transfusion since day 80 (when his hemoglobin level kept > 12.5 g/L). His beta globin gene mutation and HLA typing were all the same as the donor's analyzed on day 60 and 200. There was also a switch in blood group from A pre-transplant to O post-transplant. It is concluded that the new conditioning and GVHD prophylaxis regimens allow a successful engraftment in this case. This observation may contribute in developing UCBT as an alternative when matched sibling donors are not available.
Child, Preschool ; Cord Blood Stem Cell Transplantation ; adverse effects ; Globins ; genetics ; Graft vs Host Disease ; etiology ; HLA Antigens ; immunology ; Histocompatibility ; genetics ; immunology ; Histocompatibility Testing ; methods ; Humans ; Male ; Mutation ; Siblings ; Transplantation Tolerance ; immunology ; Transplantation, Homologous ; beta-Thalassemia ; therapy

OBJECTIVETo study the effect of selective moderate head cooling therapy on maximum length sequences brainstem auditory evoked potential (MLS-BAEP) in newborn piglets with hypoxic-ischemic brain damage.

METHODSSixteen newborn piglets aged 5-7 day old were randomly divided into three groups: normothermic control (n=4), HI (n=6) and mild hypothermia-treated (n=6). HI was induced through temporary occlusion of both carotid arteries, followed by mechanical ventilation with low concentration of oxygen (FiO2=0.06) for 30 minutes. Mild hypothermia was induced by equipment via circulating water. MLS-BAER was recorded before HI and at 12 hours, 24 hours, 36 hours, 48 hours, 60 hours, 72 hours, 4 days, 7 days, 10 days, 13 days and 15 days after HI.

RESULTSCompared with the normothermic control group, all latencies and intervals tended to increase significantly at 72 hours in the HI group and reached peak values on day 7. From day 10, all latencies and intervals tended to decrease, but apart from wave I latency, still differed significantly from those of the normothermic control group. MLS-BAER variables did not reach normal values until day 15. Ⅲ latency, Ⅰ-Ⅲ interval and Ⅰ-Ⅴ interval were significantly reduced in the hypothermia-treated group between 60 and 7 days after HI compared with the HI group (P<0.05). V latency and Ⅲ-Ⅴ interval in the hypothermia-treated group were also reduced compared with the HI group between 72 hours and 7 days after HI (P<0.05).

CONCLUSIONSBoth peripheral and central auditory systems are disturbed by HI, which shows as a significant increase in MLS-BAER variables (all latencies and intervals) in newborn piglets. Involvement in central brainstem auditory system reaches a peak on day 7 after injury. MLS-BAER variables still cannot reach to normal values until day 15. Selective moderate head cooling therapy can significantly reduce brainstem damage induced by HI.

Animals ; Animals, Newborn ; Evoked Potentials, Auditory, Brain Stem ; Hypothermia, Induced ; Hypoxia, Brain ; physiopathology ; therapy ; Swine

Objective To study the decomposition kinetics of omethoate in blood.Methods The acetonitrile precipitated protein was added into the blood, with the chromatographic column of a Waters BEH C18column (2.1 mm×50 mm, 1.7μm), the mobile phase of 5 mmol/L ammonium acetate aqueous solution-methanol, and the gradient elution with a flow rate of 0.3 mL/min and injection volume of 2μL.With electrospray ionization (ESI) source and positive ion detection, qualitative and quantitative analyses were taken using multi-reaction monitoring mode.Omethoate standard was added into blank human blood to the mass concentrations of 0.78, 1.40, 2.30, 4.50, and 7.20μg/mL, and each mass concentration was preserved at 3 temperatures of-20℃, 4℃, and 20℃, respectively.The content of omethoate was detected at different time points (0, 1, 3, 4, 7, 11, 15, 24, 32, 40, 48, 64, 80, 96, and 120 d).Results Different concentrations of omethoate all showed a descended trend in human blood under different temperature conditions.The decomposition in storage environment of-20℃, 4℃, and 20℃was fit to a one-compartment open model with a first-order kinetic process, which could be expressed as Ct=Coe-αt, with the calculated theoretical values of omethoate concentration close to the measured values.Conclusion All concentrations of omethoate are decomposed in the blood, which vary a lot in different preservation conditions.It is suggested that blood samples should be frozen and detected timely in suspected omethoate poisoning cases.

BACKGROUNDThe incidence of no reflow phenomenon limits the clinical outcomes of percutaneous coronary intervention (PCI). This randomized controlled study was designed to evaluate the immediate protective effects of intensive statin pretreatment on myocardial perfusion and myocardial ischemic injury during PCI.

METHODSAltogether 228 patients with acute coronary syndrome (ACS) were randomly assigned to standard statin group (SS group, n = 115) and intensive statin group (IS group, n = 113). Patients in the SS group received 20 mg simvastatin and patients in the IS group received 80 mg simvastatin for 7 days before PCI. Thrombolysis in myocardial infarction (TIMI) flow grade (TFG), corrected TIMI frame count (CTFC) and TIMI myocardial perfusion grade (TMPG) of the intervened vessel were recorded before and after stent deployment. Creatine kinase (CK) isoenzyme MB, troponin I and plasma level of high sensitive-C reactive protein (hs-CRP), P-selectin and intercellular adhesion molecule (ICAM) were measured before and 24 hours after the procedure.

RESULTSThe TFG after stent deployment was significantly improved with less TIMI 0-1 and more TIMI 3 blood flow in the IS group than in the SS group (all P < 0.05). Patients with no reflow phenomenon were less in the IS group (P < 0.001). The CTFC was lower in the IS group than in the SS group (P < 0.001). TMPG was also improved in the IS group than in the SS group (P = 0.001). Although PCI caused a significant increase in CK-MB 24 hours after the procedure, the elevated CK-MB value was lower in the IS group than in the SS group (18.74 +/- 8.41 vs 21.78 +/- 10.64, P = 0.018). Similar changes were also found in troponin I (0.99 +/- 1.07 in the IS group vs 1.47 +/- 1.54 in the SS group, P = 0.006). CK-MB elevation occurred in 27.8% (32/115) of the patients in the SS group vs 15.9% (18/113) in the IS group (P = 0.030). Myocardial necrosis was detected in 4.4% (5/115) of the patients in the SS group, whereas 0.9% (1/113) in the IS group (P = 0.341). But no myocardial infarction was found. Similarly, the patients with increased level of troponin I were much more in the SS group (36.5%, 42/115) than in the IS group (19.5%, 22/113) (P = 0.04). Among them, myocardial necrosis was detected in 13.0% (15/115) of the patients in the SS group, while 4.4% (5/113) in the IS group (P = 0.021). Myocardial infarction was found in 4.4% (5/115) of the patients in the SS group and 0.9% (1/113) in the IS group (P = 0.213).

CONCLUSIONSIntensive statin pretreatment for 7 days before PCI can further improve myocardial blood perfusion, protect the myocardium from ischemic injury. These effects are associated with the lowered levels of hs-CRP, P-selectin and ICAM.

Acute Coronary Syndrome ; drug therapy ; pathology ; therapy ; Aged ; Angioplasty, Balloon, Coronary ; methods ; Anticholesteremic Agents ; therapeutic use ; Female ; Heart ; drug effects ; Humans ; Male ; Middle Aged ; Myocardium ; pathology ; Simvastatin ; therapeutic use ; Treatment Outcome