The experimental study of cerebral ischemia plays an important role for understanding the pathogenesis of ischemic brain injury, but its correlation with the clinical therapeutic strategies has certain limitations. One of its main reasons is that the experimental models and methods can not or only partially repeat the pathophysiological processes of natural cerebral ischemia. In order to promote the understanding and interpretation of the experimental data, we review the commonly used experimental animal models and modeling methods and mainly elaborate the methods of current different in vivo and in vitro clinical evaluation. Based on these studies, we believe that the standardized clinical evaluations are hugely important for assessing the experimental results and clinical transformation.

Objective To investigate the diagnosis and treatment of cerebral venous sinus thrombosis ( CVST) in early pregnancy.Methods The clinical data of 4 patients with CVST in early pregnancy were analyzed retrospectively.Results The age of the 4 patients with CVST during early pregnancy was 22 -28 years old.The clinical symptom was headache, and 1 case with seizure.The clinical sign was papilledema.The MRI and MRV examination showed that the superior sagittal sinus thrombosis in 1 case, the superior sagittal sinus and left transverse sinus thrombosis in 1 case, and the left transverse sinus and sigmoid sinus thrombosis in 2 cases.The patients were received anticoagulant therapy with low molecular weight heparin, 1 case was added endovascular thrombolytic therapy.After therapy, 2 cases were cured, and 2 cases improved.Conclusions CVST in early pregnancy is easy to be misdiagnosed.MRI and MRV are helpful to diagnose it.The mainly therapies are endovascular thrombolysis and anticoagulation therapy with low molecular weight heparin.

Objective To investigate the effect of inhibition of endoplasmic reticulum stress (ERS) in ischemic preconditioning-induced cerebral ischemia tolerance.Methods A total of 120 adult male SpragueDawley rats were randomly allocated into three groups:sham operation,global cerebral ischemic and ischemic preconditioning groups (ischemic preconditioning for 3 minutes,and global cerebral ischemia for 15 minutes after 2 days).Three time points (day 1,day 3 and day 7) were set.Sugawara method was used to observe the changes of neurological behavior in rats.TUNEL staining was used to observe the conditions of cortical neuronal apoptosis.Immunofluorescence staining and Western blot analysis were used to detect the expression levels of ERS-related protein CHOP,GRP78,and caspase-12.Results The neurological behavior score showed that the sham operation group did not have neurological deficits.Both the global cerebral ischemic group and the ischemic preconditioning group had obvious neurological deficits,and they improved gradually with the passage of time,but after modeling,the neurological scores at each time point in the global cerebral ischemic were significantly lower than those in the ischemic preconditioning group:at day 1∶11.00 ±0.63 vs.14.33 ±0.33 (t =21.74,P＝0.001); at day 3∶ 12.17±0.31vs.15.17±0.48 (t=27.93,P =0.000); at day 7:14.67±0.49 vs.16.33 ±0.33 (t =7.81,P=0.020).TUNEL staining showed that at day 7 after ischemia,the positive cell count per mm2 in the sham operation,global cerebral ischemic and ischemic preconditioning groups were 4.83 ±1.85vs.395.67± 43.43 and 146.17± 27.38 respectively (F=23.62,P=0.001).The ischemic preconditioning group was significantly lower than that in the global cerebral ischemic group (P =0.001).Immunofluorescence staining showed that at day 7 after ischemia,the numbers of positive cells of CHOP (26.50±3.89vs.82.33±4.25; P=0.000),GRP78 (15.00±2.02vs.35.67±2.99; t=0.000),and caspase-12 (22.33 ± 2.76 vs.66.50± 7.25; P=0.000) in the ischemic preconditioning group were significantly less than those in the global cerebral ischemic group.Western blotting showed that at day 7 after ischemia,the expression levels of CHOP (1.22 ± 0.38 vs.3.22 ± 0.51; t =24.50,P =0.001),GRP78 (1.78 ± 0.45 vs.3.16 ± 0.76; t =14.29,P =0.005),and caspase-12 (2.89 ± 0.53 vs.5.96 ± 0.67; t =77.73; P =0.000) in the ischemic preconditioning group were significantly lower than those in the global cerebral ischemic group.Conclusions Ischemic preconditioning demonstrated a neuroprotective effect for the second lethal ischemia,its mechanism may be associated with the relief of ERS and downregulation of ERS-related protein.

Objective To investigate the change on monoamine neurotransmitter in cerebral cortex motor area of traumatic asphyxia canines and provide scientific basis for its therapy.Methods The model of canine traumatic asphyxia was established,the change on monoamine neurotransmitter in cerebral cortex (motor area) and the products of metabolism during different time were tested with HPLC DC method.Results At 2 h after damage in cerebral cortex 5 hydroxyindolecetic acid (5 HIAA) elevated remarkably; At 8 h after da mage 5 hydroxytryptamine (5 HT), homovanillic acid (HVA) elevated; but there was no obvious change on norepinephrine(NE) and 3,4 dihydroxyphenyl acetic acid (DOPAC).Conclusion The monoamine neurotransmitters might play an important role in the pathological course of secondary brain injury after traumatic asphyxia.The utilization of 5 HT antagonists or compound inhibitor at early stage was a reliable method for treating brain injury after traumatic asphyxia.

Objective To investigate costimulatory molecules CD80 and CD86 expression in acute myelogenous leukemic cells and clinical implication.MethodsThe expression of CD80 and CD86 in the patients with acute myelogenous leukemia and HL-60 cells,U937 cells,NB4 cells,K567 cells was confirmed by Flow Cytometer.ResultsCD80 was very low or no expression in patients with acute myelogenous leukemia.CD86 expressed in acute myelogenous leukemia (27.86±19.65)%,which was much higher than that in control group[(1.21±0.13)%,t＝3.55,P＜0.01].No significant changes were observed in the expression of CD86 in M4 cells(48.65±21.92)%,M5 cells(39.25±18.67)% and control group(50.20±20.31)%(P＞0.05).After the cells were cultured for 24 h and 48 h,the expression of CD86 was (30.62±5.35)% and (29.43±4.67)% in HL-60 cells ,(24.12±5.23)% and (26.56±6.54)% in U937 cells,and,(21.25±3.78)% and (23.21±6.98)% in NB4 cells (all P＞0.05).The expression of CD80 and CD86 was very low in K562 cells.ConclusionsCostimulatory molecules CD86 expressed in acute myelogenous leukemic cells in the patients with acute myelogenous leukemia and HL-60 cells,U937 cells,NB4 cells.

Objective To investigate the effects of acidosis on the current change of transient receptor potential channel M7 (TRPM7) and collagen production in mouse cardiac fibroblast (MCFs), and to explore the pathophysiological function of TRPM7 on the cardiac fibrosis. Methods (1) The model of MCFs was established and isolated. (2) MCFs was subcuhured. (3) Patch clamp technique was used to observe the current characteristics of TRPM7 in low PH solutions. (4) The influence of acidic condition on Ca2+ influx in MCFs was recorded by calcium fluorescent indicators. Results (1) There was a high level expression of TRPM7 in MCFs and the electrophysiological characteristics of TRPM7-like (TRPM7L) was similar to that of TRPM7. (2) Ca2+ influx was increased in acidic condition, and the F340/F380 ratio was increased from 1.0 to 4. 6 at pH 4.0 compared with pH 7.0 (t=2.72, P<0.01). Conclusions (1) TRPM7 is the molecular basis of the native TRPM7L in MCFs and TRPM7L plays an important role in Ca2+ influx. (2) The pathophysiological function of MCFs is influenced by regulation of Ca2+ influx mediated by TRPM7L in the condition of acidosis.

Objective To investigate the changes of TRPM 7-like current in mouse cardiac fibroblast(CFs) after myocardial infarction and the effect of myocardial ischemia on the TRPM 7 expression and current. Methods (1) The model of myocardial infarction was made and CFs were isolated;(2) CFs were cultured and infected by TRPM 7 siRNA;(3) The effects of myocardial ischemia on TRPM 7 current were recorded by whole cell patch clamp technique;(4) The influences of myocardial isehemia on Ca2+ influx in CFs were recorded by Ca2+ fluorescence imaging. (5) The effects of ischemia on total collagen content of CFs were studied. Results (1) Ca2+ inward current of CFs was increased after myocardial infarction [(16.2±1.7) vs. (7.4±0.7) pA/pF, P<0.053];(2) TRPM 7 current was reduced by 90% after siRNA infection;(3) The total collagen content of CFs after ischemia was approximately 2.3-fold higher than basic value. Conclusions Ca2+ influx in CFs plays an important role in the pathophysiological process of myocardial fibrosis.

Objective To study the role of Vibrio vulnificus cytolysin(rVvhA) induced THP-1 apoptosis and calcium influx.Methods CCK-8 cell proliferation kit,Fluo3/AM staining and AnnexinV/PI staining were performed to identify the apoptosis and calcium influx induced by rVvhA in THP-1 cells.Results rVvhA could induce THP-1 apoptosis and up-regulate the cellular calcium concentration.BAPTAAM could enhance the calcium influx induced by rVvhA in THP-1.Conclusion rVvhA had cytotoxic to THP-1 cells by inducing apoptosis and triggering extracellular calcium influx.

Objective To investigate the neuroprotective effect of sophocarpine against transient focal cerebral ischemia via down-regulation of the acid-sensing ion channel 1(ASICl) in rats.Methods Twenty-five SD rats were randomly allocated into sham operation,cerebral ischemia/reperfusion,and 5,10,and 20 mg/kg sophocarpine pretreatment groups (n=5 in each group).A rat focal ischemia model was induced by the intraluminal suture method.Five,10 and 20 mg/kg sophocarpine were injected intraperitoneally for pretreatrnent.2,3,5-triphenyltetrazolium chloride staining was used to detect cerebral infarct volume.TUNEL staining was used to detect apoptosis.Immunohistochemistry and Western blot were used to detect the expression of ASIC1 and ASIC2.Results The infarct volume after ischemia-reperfusion was(181.21±9.21)mm3,while the 5,10,and 20 mg/kg sophocarpine pretreatment groups were(150.12±6.19),(52.31±4.20),and(32.18±3.82)mm3,respectively;the neurological function scores in the cerebral ischemia/reperfusion group was(3.62±0.36),while the 5,10,and 20 mg/kg sophocarpine pretreatment grows were(3.15±0.36),(1.92±0.18),and(1.85±0.21),respectively;The surviving neurons only accounted for(31.2±2.8)% of the total cell number in the cerebral ischemia-reperfusion group,while they accounted for(51.2±3.7)%,(76.5±2.1)%,and(77.1±4.1)% in the 5,10,and 20 mg/kg sophocarpine pretreatnmat groups.Compared with the cerebral ischemia/reperfusion group,the cerebral infarct volume was decreased significantly in the sophocarpine pretreatrnent groups(all P<0.01),the neurological function scores were decreased significantly(all P<0.01),and the number of apoptotic cells was decreased significantly (all P<0.01).Immunohistochemistry showed that the number of ASIC-1 positive cells in the sham operation,cerebral ischemia-reperfusion,and 5,10,and 20 mg/kg sophocarpine pretreatment groups were(162.5±8.3),(165.1±5.3),(138.3±7.2),(82.1±6.3),and(69.2±5.5)/mm respectively;Western blot showed that the ASIC1 protein expression was decreased sigaificantly in the 10 and 20 mg/ky sophocarpine pretreatment groups (P<0.01),while there WaS no significant difference in the ASIC2 protein expression.Condusions Sophocarpine may play a neuroprotective role for cerebral ischemia-reperfusion injury in rats via down-regulating the expression of ASIC1 protein.

Objective To explore the effect of different positive end expiratory pressures (PEEP) on oxygen delivery (DO2) after recruitment maneuvers (RM) in dogs with acute respiratory distress syndrome (ARDS). Method After ARDS models established in 15 dogs by oleic acid, static P-V (pressure-volume) curves were determined by low flow technique. Lower inflection point (LIP) was set by two-way linear regression methods. RM was operated with the pressure control method. ARDS dog models were randomly divided into three groups, namely PEEP 8 cmH2O group (group A), 12 cmH2O group (group B) and 16cmH2O group (group C) after RM (equivalent to pressure at4 cmH2O under LIP, 4 cmH2O near LIP, and 4 cmH2O above LIP, respectively). Hemodynamics and arterial blood gas analysis were monitored before RM, 0, 5, 10 and 15 min after RM. The recruited volume was measured by P-V curve method 15 min after RM and respiratory mechanics was also observed at the same time. Then DO2 was calculated. The quantitative variables were summarized as the mean and SD. The t-test was used to compare continuous variables between the two independent samples. One-way analysis of variance was used to compare variables among three groups. The level of significance was set at P＜0.05 for all the tests. Results In group A, the levels of PaO2 were significantly reduced 5 min, 10 min and 15 min after RM[(257 ± 23 )mmHg, (253±21)mmHg, and (255±19)mmHg] compared with PaO2 at 0 min [(322 ± 20) mmHg] (P＜0.05).But in group B and group C, the levels of PaO2 5 min, 10 min and 15 min after RM were not lower than level of PaO2 at 0 min after RM (P＞0.05 ). The levels of PaO2 in groups B and C were higher than that in group A at the same time (P＜0.05). The recruited volume distinctly increased with PEEP levels escalated [(50±12 ) mL, (124 ±15) mL, and ( 157 ±10)mL](P＜0.05). However, the increment in the recruited volume from PEEP 8 cmH2O to 12 cmH2O was dramatically greater than that from PEEP 12 cmH2O to 16 cmH2O.There was no significant difference in static compliance between group A and B [(14.3 ± 2.2) mL/cmH2O vs. (17.2±1.4)mL/cmH2O] (P ＞ 0.05 ). But compared with groups A and B, the static compliance in the group C significantly reduced(10.5 ± 0.9) mL/cmH2O ( P ＜ 0.05 ). The ratios of DO2 after RM to DO2 before RM were different at different levels of PEEP. The levels of DO2 after RM[( 1.15 ± 0. 11 ),( 1. 14 ± 0.12), ( 1.14 ± 0. 12) and ( 1.16 ± 0.11 )] increased more greatly than that before RM ( 1.00 ±0.09) in the group B (P ＜ 0.05 ). It did not occurred in the groups A and C. Conclusions The PEEP 12 cmH2O set at near the LIP after RM could be the optimal PEEP. Not only can it improve DO2 and the static compliance, but also maintain oxygenation and the recruited volume after RM.