Objective To investigate the expression of CD8 and CD28 lymphocytes in peripheral blood of patients with breast cancer before and after operation and explore its clinical significance.Method Flow cytometry was applied to test the expression of CD8,CD8+CD28+and D8+CD28-in the peripheral blood of both 40 patients with breast cancer and 25 healthy controls. Results The expression levels of CD8 in patients with breast cancer were significantly higher than those in healthy controls [(30.4±3.3)%VS. 30.4±3.3)%,P<0.05] . The expression levels of CD8+CD28+in patients with breast cancer were significantly lower than those in healthy controls [(10.4 ± 1.7)%VS. (14.3±2.6)%,P<0.05] . The expression levels of CD8+CD28-in patients with breast cancer were significantly higher than those in healthy controls [(25.1 ±2.8)% VS. (19.6 ±2.3)%, P<0.05] . After operation, the expression levels of CD8, CD8+CD28+and CD8+CD28-[(31.9 ±4.3)%, (15.4 ±2.7)%and (20.4±2.8)%, respectively] in peripheral blood of patients with breast cancer had no significant change compared with the healthy controls. The expression levels of CD8, CD8+CD28+and CD8+CD28-in peripheral blood of patients with breast cancer had significant difference between before and after operation. Conclusions After surgical operation, the immune defence and immune surveillance of cytotoxic T lymphocytes can be enhanced, immune suppression and tumor escape can be reduced. The assays of cell phenotypes CD8 and CD28 might be useful for evaluating the immunal state and prognosis of patient s with breast cancer.

Objective To evaluate the diagnostic accuracy of diffusion tensor imaging (DTI) of corticospinal tract in amyotrophic lateral sclerosis (ALS) and find optimal testing strategies and optimal cutoff values of DTI indices for individual patient discrimination.Methods Thirty-three ALS patients and 34 healthy controls,collected at Peking Union Medical College Hospital from June 2004 through July 2005,undergoing brain DTI studies and fractional anisotropy (FA) examinations along the corticospinal tract,were analyzed by receiver operating characteristic (ROC) curves.Results Compared with the controls,ALS group had significantly decreased FA values in subcortical white matter of the precentral gyrus,the posterior limb of the internal capsule and the cerebral peduncle.In ROC analysis,the average FA value of the former two positions showed the best performance with an area under the curve of 0.917,an optimal cut-off value of 0.604,a sensitivity of 0.759 and a specificity of 0.912.The corresponding data for the average FA of all the three positions and each single position were listed as follows:average of three 0.914,0.648,0.759,0.912; precentral gyrus 0.875,0.509,0.733,0.824; internal capsule 0.845,0.692,0.656,0.941 ; and cerebral peduncle 0.752,0.742,0.656,0.735.Conclusions FA values of the corticospinal tract have a good accuracy in detecting upper motor neuron involvement in ALS.Precentral gyrus and posterior limb of the internal capsule and the average FA values of these two positions were suggested as the preferred testing places and DTI indices for clinical use.

Objective To investigate the diffusion tensor imaging(DTI)and magnetic resonance spectroscopy(MRS)changes in amyotrophic lateral sclerosis(ALS)and to study on the physiopathology of ALS.Methods Thirty-eight AIS patients,8 patients with pure lower motor neuron involvement and 5 patients with mixed-type cervical spondylosis and 34 healthy controls were enrolled and examined with the conventional brain MRI,DTI and 1H-MRS.Results T2-weighted and T2 FLAIR images revealed abnormal high signals in bilateral pyramidal tract positions in 21%(8/38)of ALS patients,which was higher than that in the coaex When compared with the control group,the values of FA in the precentral gyms(0.492±0.059 vs 0.552±0.045,F=17.150,P<0.01),the posterior limb of the internal capsule(0.679±0.048 vs 0.727 4-0.031.F=19.481.P<0.01)and the cerebral peduncle(1eft:0.740 4-0.038 vs 0.761 4±0.024.F=4.290;right:0.720 ±0.044 vs 0.746 ±0.034,F=3.264,both P<0.01)of the ALS group were decreased significantly.so were the values of NAA/Cr.And the FA reduction rate was 10.9%,6.6% and 2.8%-3.5% respectively.There was also a significant decrease in FA in the anterior limb of the internal capsule and the occipital lobe in the ALS group.Conclusion Not only does the application of DTI and MRS help to demonstrate upper motor neuron involvement in the brain and facilitate the diagnosis and differential diagnosis of ALS,but also increases the understanding of the lesion distribution and physiopathology in ALS.

0.05), CGI and HAMA. Safety analysis suggested that no significant differences were found in symptoms and frequency of side effects between the two groups.Conclusion Bupropion hydrochloride tablet is an effective and safe antidepressant. It has similar effect and safety compared with fluoxetine in the treatment of depression.

We conducted purification of filamentous temperature-sensitive protein Z of Streptococcus suis serotype 2 (S.suis 2) and measured its GTPase activity.The ftsz gene in the genome of the Chinese 05ZYH33 strain of S.suis 2 was successfully amplified using PCR,and then the ftsz gene was cloned into prokaryotic expression plasmid pET28a,and the recombinant plasmid pET28a-ftsz was transformed into E.coli BL21.After induction by IPTG,the isolated FtsZ protein was analyzed with SDS-PAGE.Then the recombinant protein was purified by Ni2+-NTA affinity chromatography.The rabbit serum was harvested after immunization with recombinant FtsZ protein,and was analyzed by indirect ELISA and Western blotting.The GTPase activity of FtsZ was measured with the malachite green method.Results showed that successfully constructed recombinant plasmid pET28a-ftsz and the recombinant protein with high purity was obtained;Western blot result indicated that FtsZ could react with the His-tag antibody and the rabbit serum;the polyclonal antibody titer of the rabbit serum reached 1 ∶ 13 107 200;FtsZ have GTPase activity.We successfully prepared S.suis 2 recombinant protein FtsZ having GTPase activity and high titer antiserum would be useful for the further study of S.suis 2 cell division mechanism.

Objective To investigate the effects of CD24 on CCl4-induced murine liver fibrosis and to analyze the possible molecular mechanism.Methods Wild type (WT) and CD24 knockout (CD24-/-) C57BL/6 mice were treated with CCl4 through intraperitoneal injection.Levels of ALT in serum samples were detected and liver tissues were stained with hematoxylin and eosin (HE) to assess liver tissue injury.Sirius Red staining was used to observe liver fibrosis.Real-time PCR was performed to detect the expression of α-SMA (α-smooth muscle actin), Col1a1 (Collagen, typeⅠ, alpha 1), TGF-β1 (transforming growth factor-β1) and CD24 at mRNA level in liver tissues.Western blot was performed to analyze the expression of α-SMA and Col1a1 at protein level.Flow cytometry analysis was used to detect the macrophages in liver tissues.ELISA was used to detect the expression of TGF-β1 in the culture supernatants of M1 and M2 macrophages.Results The expression of CD24 at both mRNA and protein levels were up-regulated in mice with CCl4-induced liver fibrosis.HE staining showed that liver inflammation in CD24-/-mice was more severe than that in WT mice after treated with CCl4.Sirius Red staining of paraffin-embadded liver sections revealed that compared with WT mice, CD24-/-mice presented with more severe liver fibrosis.Moreover, α-SMA and Col1a1, indicators of liver fibrosis, in CD24-/-mice were significantly higher than those in WT mice.Flow cytometry analysis showed that murine hepatic macrophages significantly increased in CD24-/-mice than in WT mice following CCl4 treatment.Real-time PCR analysis also confirmed that significantly enhanced expression of TGF-β1 at mRNA level in liver tissues was observed in CD24-/-mice than in WT mice.TGF-β1 secreted in the culture supernatant of M2 macrophages derived from CD24-/-mice group was more than that of the WT mice group.No significant difference in TGF-β1 secretion in culture supernatant of M1 macrophages was observed between the two groups.Conclusion Taken together, these data suggest that CD24 plays an important role in attenuating CCl4-induced chronic inflammation and hepatic fibrosis in mice.The mechanism of CD24 in alleviating liver fibrosis might be through regulating intrahepatic macrophages, inhibiting the secretion of TGF-β1 by M2 macrophages and suppressing the activation of hepatic stellate cells.

Although myocardial ischemic condition can be improved by reperfusion therapy, the sequent myocardial ischemiareperfusion injury may associate with the adverse outcomes in patients with ST-segment elevation myocardial infarction. In the recent years, several therapies, such as ischemia preconditioning and ischemia postconditioning, have been proven effective in animal experiments and have been widely incorporated into clinical trials for the reduction of ischemia/reperfusion injury. Based on the previous clinical evidences, we review the study progress of ischemia conditioning in patients with ST-segment elevation myocardial infarction for optimizing the management of acute myocardial infarction.

In order to screen the inhibitors of GAT1 protein, the effects of synthetic N-acylglucosamines on the GABA uptake activity of GAT1 were examined in the HEK-293 cell model stably expressing GAT1 by performing GABA uptake assay. And since the N-linked oligosaccharides, especially their terminal sialic acid of GAT1 are necessary for the function of GAT1, their effects on the modification of N-glycans of GAT1 were also determined by the quantitative analysis of the sialic acid of GAT1. The results showed that 10 mmol /L 3-O-methyl-N-acetylglucosamine(3-O-Met-GlcNAc)reduced the GABA uptake activity of GAT1 to 53%(P< 0. 01)through inhibiting the sialylation of GAT1(66. 8%). And the GABA uptake activities were decreased to 54%, 63%, 63% and 67%(P< 0. 01), respectively, by the treatments of 10 mmol /L N-propionylglucosamine(GlcNProp), N-hexanoylglucosamine(GlcNHex), N-cycloproylgormylglucosamine(GlcNCyclo)and N-acetamidoacetylglucosamine(GlcNAc-acetamido)through inhibiting N-glycan trimming. These results indicate that the analogues of glucosamines have great potential in the development of inhibitors of GAT1 activity or sialic acid biosynthesis.

ObjectiveTo study the effect of pre-arrest and post-arrest mild hypothermia after restoration of spontaneous circulation (ROSC) on myocardial function, ultrastructure, apoptosis of myocardial cells in rabbits with ventricular fibrillation.Methods Sixty-two male New Zealand rabbits were randomly allocated into five groups: namely normothermic control group (NTC group,n = 10), hypothermia control group (HTC group,n = 10), normothermic resuscitation group (NTR group,n = 14), hypothermia pre-arrest group (HPRA group,n = 14), and hypothermia post-arrest group (HPOA group,n = 14). The normal temperature was controlled at (39.0±0.5)℃, and the hypothermia (33.5±0.5)℃. Ventricular fibrillation cardiac arrest (CA) was reproduced in rabbits by transcutaneous epicardium electrical stimulation. The parameters of hemodynamics were monitored dynamically for 4 hours in all the groups, including heart rate (HR), left ventricular end diastolic and systolic pressure (LVEDP/LVESP), maximal rate of increase/decrease in left ventricular pressure (±dp/dt max), and mean arterial pressure (MAP). The body temperature of rabbits in hypothermia groups was maintained by surface cooling for 4 hours followed by rewarming. The survived rabbits were sacrificed at 48 hours after resuscitation, and myocardial apical tissue was harvested for observation of ultrastructure with electronic microscope, and to observe apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining.Results① Resuscitation investigation: there was no significant difference in rate of ROSC, time of CPR and energy of defibrillation among HPRA, HPOA, and NTR groups [rate of ROSC: 85.71%, 71.43%, 71.43%; time of CPR (seconds): 45.3±30.2, 61.2±41.3, 82.3±63.8;energy of defibrillation (J): 14.3±8.9, 22.0±15.5, 25.0±15.8, allP> 0.05].② Hemodynamics: compared with normal temperature groups, animals in hypothermia groups exhibited lower levels of HR (allP< 0.05). Compared with NTR group, HPRA group exhibited higher levels of LVESP (mmHg, 1 mmHg = 0.133 kPa) at 0.5, 1, 2 and 3 hours post ROSC (0.5 hour: 103.8±14.3 vs. 91.6±13.3, 1 hour: 107.2±14.1 vs. 82.7±8.5, 2 hours: 109.0±16.9 vs. 88.8±12.9, 3 hours: 109.1±14.6 vs. 89.3±14.3, allP< 0.05). Compared with NTR group and HPOA group, HPRA group exhibited lower levels of LVEDP (mmHg) at 0.5 hour post ROSC (3.70±0.85 vs. 7.61±2.73, 7.02±3.12, both P< 0.05). Compared with NTR group, HPRA group exhibited lower levels of LVEDP at 1 hour post ROSC (4.34±1.44 vs. 6.99±1.96,P< 0.05). In HPRA group, the level of+dp/dt max (mmHg/s) was higher than that of NTR group and HPOA group at 1 hour and 2 hours post ROSC (1 hour: 2 759.5±321.6 vs. 2 123.0±304.5, 2 283.7±234.2, 2 hours:2 730.6±425.1 vs. 2 221.5±392.9, 2 252.6±476.0, allP< 0.05). There were no significant differences in -dp/dt max and MAP levels among three CPR groups.③ The survival rate at 48 hours post ROSC of NTR, HPRA and HPOA groups was 60%, 75%, and 100%, respectively. Compared with NTR group, higher survival rate was found in HPOA group at 48 hour post ROSC (P< 0.05).④ Compared with NTR group, less damage to myocardial ultrastructure was found in HPRA and HPOA groups. Apoptosis index (AI) was lower in HPRA and HPOA groups than that in NTR group [(28.05±9.82) %, (26.39±8.98) % vs. (42.02±13.36) %, bothP< 0.05].Conclusions Our study shows that mild hypothermia has no effect on ROSC rate. Pre-arrest hypothermia can ameliorate myocardial systolic function of rabbit in early stage after ROSC, and it has no negative influence on diastolic function. Post-arrest mild hypothermia produces no negative influence on myocardial function of rabbit, but it improves 48 hours survival rate in ROSC rabbits. Both pre-arrest and post-arrest mild hypothermia therapy can attenuate myocardial injury in CA model of rabbits by ameliorating mitochondrial injuries and suppressing apoptosis of myocardial cells.

Objective To compare the effects of airway humidification by intermittent atomization inhalation and micro pump control on pulmonary infection after tracheotomy in elderly patients with serebral hemorrhage. Methods From February 2012 to February 2013, 46 elderly patients with heavy craniocerebral injury and tracheotomy were set as the control group, treated with intermittent atomization inhalation. From March 2013 to March 2014, 50 elderly patients with severe craniocerebral injury and tracheotomy were set as the observation group, treated with micro pump controlled continuous airway humidification. The two groups were compared in terms of the satisfaction rate and incidence of pulmonary infection. Result The satisfaction rate of the observation group was higher than that of the control group and the incidence of pulmonary infection was significantly lower than that of the control group (P<0.05). Conclusion Micro pump controlled airway humidification for elderly patients with severe craniocerebral injury can improve the airway humidification effect and reduce the incidence of pulmonary infection.