Objective Effective tissue perfusion fixation plays an important role in immunofluorescence experiments and other studies .The aim of this study was to modify the method for perfusion fixation of rat brain tissues and improve the efficiency of the procedure . Methods Conventional thoracotomy was modified by cutting the skin from the xiphoid along the mediventral line to the lower jaw to expose both sides of the thorax and then freeing the chest wall .The tip of the infusion needle was cut off and the needle in-serted into the left ventricle , pushed to the ascending aorta , and fixed there to prevent sliding .Perfusion and fixation began after tho-racic aortic occlusion.The regulatory valve was opened , followed by infusion of 0.9%physiological saline and cutting the right atrial appendage .When the perfusate became clear , the infusion tube was replaced , without removal of the needle , and fixed with 4%paraformaldehyde .The fixation process was slowed down gradually till the neck became stiff , which indicated its completion . Results The head, neck and upper limbs were all fixed desirably by this method , which shortened the operation time , saved 4%paraformalde-hyde, and achieved satisfactory immunofluorescence results as compared with conventional procedures . Conclusion The modified method is simple , obviously improves the results of perfusion fixation of rat brain tissue , and therefore deserves to be popularized .

Objective To evaluate the effect of Shenfu injection(参附注射液) on hemodynamics and oxygen delivery(DO2) metabolism in dogs with cardiogenic shock.Methods A dog model of acute myocardial infarction was made by ligating the midpoint of left coronary artery.When the cardiogenic shock was found by the Swan-Gans vessel,18 dogs with cardiogenic shock were divided randomly into three groups: Shenfu,dopamine and control groups.The following drugs were intravenously administered: Shenfu injection 1 ml/kg to the Shenfu group,dopamine 10 ?g?kg-1?min-1 to the dopamine group and NaCl 5 ml/kg to the control group.Before the model was made and after the drugs were given for 30,60,90,120 and 180 minutes,the changes of the parameters of hemodynamics were monitored through the Swan-Gans vessel,inculding cardiac output(CO),pulmonary arterial wedge pressure(PAWP), pulmonary artery pressure(PAP),central venous pressure(CVP),heart rate(HR),blood pressure(BP) and angiosthenia(AP).Then the results of stroke volume(SV),stroke work(SW),mean arterial blood pressure(MAP),systemic vascular resistance(SVR) and pulmonary circulation resistance(PVR) were calculated by formulae.Arterial blood and mixed venous blood were used for blood gas analysis to measure and calculate the following hemodynamic parameters: DO2,oxygen consumption(VO2) and oxygen uptake rate(ERO2).The results were compared.Results ① In the control group,after NaCl was given,CO,SV,SW,HR and MAP were decreased gradually at the time points,while the PVR,PAWP and CVP were progressively increased at the same time(all P

Objective To investigate the relationship between primary TCM syndromes and heart rate variability in hypertension, evaluate the development of disease. Methods 182 hypertension patients were recruited in accordance with the standard of hypertension. According to the Chinese medicine dialectical criteria, these patients were further divided into 5 groups, namely, hyperactivity of liver-fire group, phlegm-dampness retention group, blood stasis group, hyperactivity of Yang due to yin deficiency group and Qi and blood deficiency group. Another 60 healthy people were recruited as a control group. 24h Holter was adopted to monitor heart rate variability. Results Analysis of TCM syndromes and heart rate variability, anger on inflammation group had positive relation with SDNN, RMSSD, LF, HF and TP(r were respectively 0.161,0.372, 0.223, 0.281, 0.301);Phlegmy wet resistance groups had positive relation with SDNN, HRV triangular index, VLF, LF, VLF/HF, LF/HF, LFnorm.(r were respectively 0.175, 0.211, 0.174, 0.262, 0.159, 0.264, 0.263);blood stasis group had negative relation with LF/HF, LFnorm(r were respectively-0.185、-0.018).Yang due to yin deficiency group had negative relation with SDNN, RMSSD, VLF, LF.(r were respectively -0.204, -0.161,-0.184,-0.197);Qi and blood deficiency group had negative relation with SDNN, SDANN, RMSSD and HRV triangular index, VLF, LF, LFnorm, and had positive relation with HFnorm(r were respectively-0.353,-0.348,-0.258, -0.303, -0.268, -0.361, -0.352, 0.267). Conclusion Hypertension patients with different type of TCM syndromes had different characteristics of autonomic nerve functional disorders, which both reflected the important pathological changes of TCM syndromes and had close relation with the course of hypertension, providing objective basis for clinical diagnosis of syndrome differentiation.

Age-related cataract and age-related macular degeneration (AMD) are the leading causes of visual loss in elder persons,and sometimes the two diseases are co-existing in those patients.Earlier studies have suggested that cataract surgery could promote the process of AMD.However,the latest research of this diseases and the advanced treatment strategy have leading the remarkable change of this field.This article reviews the morbidity and the relation of cataract surgery and progression of AMD,and how to reduce the development of AMD after cataract surgery.

Objective To determine the incidence and effect of unrecognized cardiac injury in critically ill patients and evaluate the significance of elevations of serum troponin Ⅰ and APACHE Ⅱscore in patients with critically illness.Methods We measured the level of serum troponin Ⅰ and evaluated relationship between elevations of serum troponin Ⅰ and APACHE Ⅱ score,myocardial injury,mechanical ventilation,intensive care unit stay by means of retrospective chart review and analysis of clinical data.Results Thirty-four(21.4%)of the 159 patients had evidence of myocardial injury based on elevated levels of cardiac troponin Ⅰ,only 9(26.5%)of these 34 patients were diagnosed as having acute myocardial infaction by the intensive care unit staff.Mortality in patients with myocardial injury that was unrecogniazed(41.2%)or recognized(44.4%)was higher than in those without myocardial injury(16.0%)(P

Objective To investigate the effect of recruitment maneuver (RM) on acute respiratory distress syndrome (ARDS).Methods A total of 7 patients with early ARDS were involved in this investigate with pressure-control ventilation of FiO2 =1.Hemodynamics and extravascular lung water index (EVLWI) were monitored.After airway preparation,the ventilation parameters were adjusted to inspiration pressure (Pi) =24 cm H2O,positive end-expiratory pressure (PEEP) =10 cm H2O,frequency (f) =50,inspiration/expiration (I/E) =5/1.Up regulated PEEP to 24 cm H2O and Pi to 45 cm H2O for 3 respiration cycles.Then down regulated Pi to 30 cm H2O and had the artery blood gas analysis immediately.If PaO2 ＜ 450 mm Hg,Pi was increased 5 cm H2O above the last one,until PaO2 ＞ 450 mm Hg,which was considered RM success.Then PEEP was decreased 2 cm H2O and repeated the cycle as formers until PaO2 ＜350 mm Hg.The optimal PEEP was 2cm H2O above that.Maintained this airway pressure at least for 4 hours.Artery blood gas and hemodynamics of pre-and post-RM were monitored.The RM would be stopped by any unstable hemodynamic.Results In 6 of 7 patients,PaO2 and PEEP were higher than the baseline (P ＜ 0.01) after 4 hours of RM ; EVLWI decreased (P ＜ 0.01).RM was stopped in 1 patient,because of the unstable hemodynamic.None of the 7 patients got RM complications ; meanwhile,6 patients were cured.Conclusions Applying RM on ARDS is safe and effective.

ObjectiveTo explore the effects and possible mechanisms of VCR combined with low dose cyclophosphamide(CTX) intermittently to treat severe systemic lupus erythematosus(SLE).It is assumed that this might be a new combination therapy for SLE and expected to improve the overall prognosis and outcome of SLE.MethodsMurine chronic graft-versus-host disease(cGVHD) model were developed for study.They were randomly divided into the control group,vincristine (VCR) pulse therapy group,CTX pulse therapy group,CTX every other day(EOD) group,VCR+CTX combination group.One way ANOVA and repeated measure variance analysis were used for statistical analysis.Results① Six weeks after cGVHD models were set up,the average 24-hour urine protein quantification was(5.02±0.88) mg,anti-dsDNA antibody was positive,and Ⅳ LN pathology could be observed histologically in the model murine.So cGVHD models were successfully developed.② Significantly difference in decreasing of 24-hour urine protein quantification was found in the CTX EOD group,VCR+CTX combination group and other groups (P＜0.01).Significant decrease in Cr,ALT,anti-dsDNA,was found in the CTX EOD group,VCR+CTX combination group,CTX pulse therapy group and other groups(P＜0.05).Decrease in urine MCP-1 and TGF-β1 could be detected,and statistical significant difference in these parameters could be found in the CTX EOD group,CTX pulse therapy group,VCR+CTX combination group and other groups (P＜0.01).MCP-1 and TGF-β1'expression in model kidney were reduced in the CTX EOD group,VCR+CTX combination group and had statistical significant difference in the CTX EOD group,VCR+CTX combination group,VCR pulse therapy group,and CTX pulse therapy group.③ VCRand CTX combination treatment was effectivein 24-hour urine protein quantification,blood Cr,ALT,anti-dsDNA and urine MCP-1,as well as urine TGF-β1 (P＜0.05).Conclusion ① The combination of VCR and CTX is synergistic in decreasing 24-hour urine protein quantification,Cr,and the expression of MCP-1,TGF-β1.② The adverse effect of VCR+CTX combination group is similar to VCR pulse therapy group and CTX pulse therapy group.

ObjectiveTo examine whether Shenfu injection (SFI) reduces post-resuscitation myocardial dysfunction in a pig model by modulating expression imbalance of transcription factors of regulatory T cell, namely GATA-3 and T-bet.Methods Thirty pigs were randomly divided into sham group (n = 6) and cardiopulmonary resuscitation (CPR) group (n = 24) according to the random number table method, and the pigs in the CPR group were randomly subdivided into normal saline (NS) group, epinephrine (EP) group, and SFI group (n = 8 per group). After 8minutes of untreated ventricular fibrillation (VF) followed by 2 minutes of CPR, animals in three groups respectively received central venous injection of either 20 mL SFI (1.0 mL/kg, SFI group), EP (0.02 mg/kg, EP group) or NS (NS group). Blood samples were obtained before VF and 0.5, 2, 6 hours after restoration of spontaneous circulation (ROSC), and the parameters of hemodynamics and oxygen metabolism were determined. Surviving pigs were sacrificed at 24 hours after ROSC, the pathological changes in myocardium were observed, the levels of interleukin-4 (IL-4), tumor necrosis factor-α (TNF-α) andγ-interferon (IFN-γ) were measured by enzyme linked immunosorbent assay (ELISA), and expressions of protein and mRNA of GATA-3 and T-bet were determined by Western Blot and quantitative real-time polymerase chain reaction (RT-qPCR), respectively.Results Six pigs of three resuscitation groups were successfully resuscitated. The CPR time, number of defibrillation, defibrillation energy, and ROSC time were significantly decreased in the EP and SFI groups compared with those in the NS group. Compared with the sham group, the parameters of left ventricular systolic function and oxygen metabolism were significantly decreased, myofibril organelles were extensively damaged, and progressive and severe deterioration of the myocardium was found, and mitochondrial structure was not recognizable in the NS group; the level of IL-4 in myocardium were markedly decreased, while that of TNF-α, IFN-γand IFN-γ/ IL-4 [reflecting helper T cell 1/2 (Th1/Th2)] were significantly increased. Protein and mRNA expressions of GATA-3 were markedly reduced in the myocardium of pigs in the NS group compared with that of the sham group at 24 hours after ROSC, while T-bet was significantly increased. Compared with the NS group, animals treated with SFI had minimal myocardial intracellular damage, with decreased heart rate (HR, bpm: 90.33±3.79 vs. 106.83±5.36) and increased mean arterial pressure (MAP), cardiac output (CO), oxygen delivery (DO2), and oxygen consumption (VO2) at 6 hours after ROSC [MAP (mmHg, 1 mmHg = 0.133 kPa): 107.67±1.96 vs. 86.83±1.85, CO (L/min): 2.47±0.08 vs. 2.09±0.04, DO2 (mL/min): 364.31±4.21 vs. 272.33±3.29, VO2 (mL/min): 95.00±2.22 vs. 82.50±2.28, allP<0.05]. Compared with the NS groups at 24 hours after ROSC, level of IL-4 was markedly increased in myocardial cells (ng/L: 33.80±3.06 vs. 16.15±1.34,P< 0.05), while the levels of TNF-α, IFN-γ and IFN-γ/IL-4 were lowered significantly [TNF-α (ng/L): 18.16±0.71 vs. 29.64±1.89, IFN-γ (ng/L): 373.75±18.36 vs. 512.86±27.86, IFN-γ/IL-4: 16.15±1.34 vs. 33.80±3.06, allP< 0.05], and myocardial T-bet protein and mRNA expressions were reduced [T-bet protein (gray value): 0.41±0.07 vs. 0.59±0.11, T-bet mRNA (2-ΔΔCt): 4.37±0.21 vs. 7.57±0.55, bothP< 0.05], furthermore, myocardial GATA-3 protein and mRNA expressions were significantly up-regulated in SFI group [GATA-3 protein (gray value): 0.25±0.07 vs. 0.16±0.07, GATA-3 mRNA (2-ΔΔCt): 0.63±0.07 vs. 0.34±0.05, bothP< 0.05]. The parameters in SFI group were significantly improved compared with those of the EP group.ConclusionsMyocardial immune dysfunction is induced by Th1/Th2 imbalance following myocardial injury subsequent to CPR in pigs. SFI can attenuate myocardial injury and regulate myocardial immune disorders, protect post-resuscitation myocardial injury by modulating expression imbalance of transcription factors GATA-3 and T-bet.

Objective To observe the effect of paclitaxel long-circulating thermo-sensitive liposome ( PLTL) on inhibiting the growth of transplanting Lewis lung cancer cells in C57BL/ 6 mice. Methods The model of mice carried Lewis lung cancer was established. 40 tumor-bearing mice were divided into five groups randomly: blank control group, model control group, PTX group, PTL group and PLTL group, 8 mice for each group. The blank control group and the model control group were injected with 0. 9% sodium chloride solution. In PTX group, PTL group and PLTL group, the dose of per injection was calculated with the reference of PTX for 20 mg·kg-1 , diluted with 0. 9% sodium chloride solution, and the mice were injected via the tail vein with a volume of 0. 2 mL each time. Except for the blank control group, 5 minutes after administration, the tumors of the other groups were subjected to local hyperthermia at (42±0. 5) ℃ for 30 min. During the treatment period, transplantation tumor growth was observed; pathological morphology changes of tumor tissues and cells were detected by HE stain; apoptosis rate of tumor cells was determined by flow cytometry to investigate the inhibition effect of PLTL combined with local thermal therapy on the tumor. Results The inhibition rate of tumor in model control group, PTX group, PTL group and PLTL group was 21. 81% , 48. 87% , 57. 22% and 78. 87% , respectively. The apoptosis rate of tumor cells was (20. 4 ± 4. 2)% , (42. 7 ± 3. 8)% , (54. 6±2. 9)% and (69. 7±5. 0)% , respectively. Observed by pathology, apoptosis rate and necrosis number of tumor cells in PLTL group were significantly increased. Conclusion Compared with PTX and PTL, PLTL has an evident thermo-sensitive feature and can increase the anticancer effect of paclitaxel remarkably when combined with local hyperthermia.

AIM:To study the effects of N-palmitoylethanolamide ( PEA) on the anxiety-and depression-like behaviors of the mouse model induced by restraint stress , and to explore the possible mechanism of anxiolytic and antide-pressant effects of PEA .METHODS:The mice were intragastrically treated with 2.5, 5 and 10 mg/kg of PEA for 21 con-secutive days once daily .Thirty min after intragastric administration , the mice ( except the normal control group ) were placed in the glass tube to accept 4-h chronic restraint stress for 21 d.After the last administration , the mice were submit-ted to the forced stress test and the open field test (OFT) to observe the effects of PEA on the depression-like behaviors. The cumulative immobility time was recorded during the 4-min interval in the forced swimming test ( FST) or during the 5-min interval in the tail suspension test (TST).The elevated plus maze (EPM) test was used to investigate the effect of PEA on the mouse anxiety-like behaviors , and the water maze method was used to investigate the learning and memory abi-lities, spatial orientation and cognitive function of mice .After the behavior tests , the serum was collected and the hippo-campus was removed . The serum contents of adrenocorticotropic hormone ( ACTH ) , cortisol ( CORT ) and 5-hydroxytryptamine (5-HT) in the hippocampus were detected by ELISA .The changes of acetylcholinesterase ( AChE) ac-tivity in the hippocampal homogenate was measured by spectrophotometry .RESULTS:Compared with model group , in the FST or TST, the immobility time in the mice treated with PEA at 2.5~10 mg/kg and fluoxetine was significantly reduced . In the OFT, the total locomotion distance and total movement time were increased significantly in the mice , but only 10 mg/kg PEA and fluoxetine increased the numbers of rearing .In the EPM test , the percentage of the time spent in open arms, the entries into open arms and the total locomotion distance in 4 arms in the mice were significantly increased .In wa-ter maze test , PEA at 5 and 10 mg/kg and fluoxetine significantly shortened the latency to find the security zone in the mice, and PEA at 10 mg/kg and fluoxetine obviously shorten the swimming distance .Compared with model group , PEA at 10 mg/kg and fluoxetine reduced the mouse serum levels of ACTH and CORT , and the adrenal index , increased the 5-HT content and decreased the AChE activity in the hippocampus .CONCLUSION:PEA produces antagonistic effects on an-xiety-and depression-like behaviors in the mice induced by restraint stress .Its specific mechanism may be related to the re-gulation of the hypothalamic-pituitary-adrenal axis function by increasing the 5-HT level in hippocampus , thus participating in the regulation of central cholinergic system .