Objective To study the relationship between serum Estradiol(E2 ) andβ-amyloid protein(β-AP) in patients with cer-ebral infarction .Methods A total of 46 female patients with cerebral infarction were selected from 2011 January to 2014 June into observation group ,46 cases were divided as 20 cases in mild group ,16 cases in moderate group ,10 cases in severe group on the basis of neural function defect score standard ,according to the size of infarct volume 46 patients were divided into small infarct volume group with 21 patients ,moderate infarct volume group with 14 cases ,larger infarction volume group with 11 cases .In the same peri-od ,64 healthy women were recruited into control group .Serum E2 ,β-AP levels of serum and cerebrospinal fluid were detected and compared ,and the relationship of serum E2 andβ-AP level in serum ,cerebrospinal fluid were analyzed .Results Theβ-AP levels in serum and cerebrospinal fluid in observation group were higher than those of control group ,the serum E2 level was significant lower than that of the control group ,the differences were significant(P<0 .05) .With more severe cerebral infarction ,β-AP levels in serum cerebrospinal fluid showed an upward trend ,with more severe cerebral infarction ,the serum level of E2 was significantly decreased , andβ-AP ,E2 levels in mild ,moderate ,severe group had significant differences(P<0 .05) .Theβ-AP levels of serum and cerebrospi-nal fluid in higher infarct group were higher than those in small ,moderate infarction group ,but E2 levels were lower than those in small ,moderate infarction group ,the differences had significant(P<0 .05) .There were no significant difference inβ-AP of serum and cerebrospinal fluid ,Serum E2 level between small and moderate infarction group(P>0 .05) .The expression of serum E2 andβ-AP in serum and cerebrospinal fluid were negatively correlated(r= -0 .428 ,P=0 .009;r= -0 .476 ,P=0 .005) .Conclusion β-AP levels in serum and cerebrospinal fluid in patients with cerebral infarction are high ,closely related with the severity of cerebral in-farction .β-AP levels of cerebrospinal fluid ,serum in cerebral infarction patients are negatively related to the level of serum E2 .

Ovarian epithelial carcinomas are the most common lethal gynecological malignancies. Ovarian carcinomas are divid-ed into Types I and II based on different morphologies, genetic alterations, and biomarker expression. Low-grade micropapillary serous carcinoma are Type I tumors. Type I tumors are slow growing, generally confined to the ovary at diagnosis, and with better prognosis. These tumors develop from well established precursor lesions that are termedborderlinetumors. Type 1 tumors are genetically stable and are characterized by mutations in a number of different genes including KRAS, BRAF, PTEN, and beta-catenin. Type II tumors are rapidly growing and highly aggressive neoplasms, for which well defined precursor lesions have not been described. They may arise in the fimbrial epithelium of the oviduct with advanced stage, more aggressive behavior, and worse prognosis. High-grade serous carcino-ma belongs to Type II tumors. This group of tumors has a high level of genetic instability and is characterized by TP53 mutation. Hence, ovarian cancer comprises a heterogeneous group of tumors with distinctly different histological characteristics, molecular genet-ic features, and clinical course.

[Objective]To explore the correlation of the expression of alpha-fetoprotein(AFP)mRNA in the peripheral blood and postoperative survival and metastasis of patients with liver cancer.[Methods] A total of 66 patients with liver cancer who received radical resection surgery from January 2005to December 2006 was enrolled in this study.The cell total RNA was extracted from peripheral blood and the expression of AFP mRNA was detected by nested PCR.All the patients were followed up for 60 months after surgery.[Results]The expression rate of AFP mRNA in the peripheral blood was 40.91％(27/66).The expression of AFP mRNA in the peripheral blood in patients with liver cancer was significantly related to microvascular invasion and metastasis(P ＜ 0.05 or ＜ 0.01),but the expression had no relationship with sex,age,HBV infection,cirrhosis,AFP concentration,tumour size and number,and Edmondson grading(P＞0.05).The overall 1,2,and ＞ 3 years survival rates of patients with positive AFP mRNA after surgery were 66.7％(18/27),38.9％(7/18),28.6％(2/7),respectively.The overall 1,2,and ≥3 years survival rates of patients with negative AFP mRNA after surgery were 84.6％(33/39),60.6％(20/33),45.0％(9/20).There was statistical significance between the survival rates of AFP mRNA-negative patients and AFP mRNApositive patients(P ＜ 0.01).[Conclusions] The detection of AFP mRNA in the peripheral blood may provide clue for early microscopic metastasis.It can be a prediction index for postoperative recurrence.

Objective To investigate the pathological role of endoplasmic reticulum stress in fluomsisinduced apoptosis in human hepatocellular carcinoma cell line (HepG2).Methods Under stimulation of 1,3,6,9 mmol/L concentrations of NaF in vitro for 24 h,while normal control group was cultured under normal condition,the apoptosis of HepG2 cells was measured by flow cytometry.The endoplasmic reticulum stress markers (glucose regulative proteins 78,94;GRP78,GRP94) and CCAAT/enhancer-binding protein homologous protein (CHOP) in HepG2 cells were measured at both mRNA and protein levels by real-time PCR and Western blotting,respectively.Results After treated with 0,1,3,6,9 mmol/L NaF for 24 h,the apoptosis rate of HepG2 cells was (6.25 ± 1.27)％,(13.48 ± 1.00)％,(24.08 ± 1.88)％,(30.19 ± 3.07)％ and (37.72 ± 4.43)％,respectively,and the difference was statistically significant among groups (F =65.828,P ＜ 0.01).After treated with 3 mmol/L NaF for 24 h,the mRNA level of GRP78,GRP94 and CHOP was (1 172.41 ± 459.60)％,(946.95 ± 635.85)％ and (7 846.97 ± 1 670.01)％,which was increased compared to those of the control groups [(100.00 ± 1.77)％,(100.00 ± 2.08)％,(100.00 ± 0.74)％,t =12.77,4.67,11.50,all P ＜ 0.01].Under the same condition,the protein levels of GRP78 and CHOP were (159.99 ± 67.59)％ and (155.15 ± 94.24)％,which were increased compared to those of the control groups [(100.00 ± 30.68)％,(100.00 ± 41.44)％,t =-3.27,-1.99,all P ＜ 0.05],while GRP94 protein level [(46.40 ± 41.46)％] was decreased compared to that of the control group [(100.00 ± 68.86)％,t =4.02,P ＜ 0.05].Conclusion Endoplasmic reticulum stress may be involved in NaF-induced cell death in HepG2 cells.

Objective To explore the potential mechanism of severe liver injury shortly after withdrawal of antiviral therapy in chronic hepatitis B (CHB) patients.Methods Forty-nine patients with chronic hepatitis B virus (HBV) infection from the Department of Infectious Diseases of the First Affiliated Hospital of Nanchang University and 8 healthy volunteers from August 2014 to March 2015 were included in this study.All of them were human leukocyte antigen (HLA)-A2-positive.CHB patients were classified into three groups,including 15 cases in immune-tolerance group,20 cases in sustained antiviral treatment group,and 14 cases in recurrence of drug withdrawal group.The frequency of peripheral HLA-A0201-restricted hepatitis B core antigen (HBcAg)18-27 pentamer complex specific CD8+ T cells in CHB patients was analyzed by flow cytometry.Enzyme linked immunospot assay(ELISPOT) was used to detect interferon-gamma (IFN-γ) and tumor necrosis factor-α (TNF-α) secretions of HBcAg18-27-specific CD8+ T cells.The experimental data were analyzed using non-parametric U tests.Results In healthy control group,immune-tolerance group,sustained antiviral treatment group and recurrence of drug withdrawal group,the frequencies of HBcAg-specific CD8+T cells were (0.17 ± 0.16) ％,(1.46±0.72)％,(3.24± 1.60)％ and (4.67±2.43)％,respectively.Compared with healthy control group,the difference were all statistically significant in the three groups (Z=-3.583,-4.018 and-3.823,respectively;all P＜0.01).The frequencies of HBcAg-specific CD8+T cells in immune tolerance group or recurrence of drug withdrawal group were both significantly different from that in sustained antiviral therapy group (Z=-3.400 and-2.030,respectively;both P＜0.05).The difference between immune-tolerance group and recurrence of drug withdrawal group was also significant (Z =-3.230,P＜0.01).The secretion levels of IFN-γ of HBcAg-specific CD8+T cells in healthy control group,immune-tolerance group,sustained antiviral treatment group and recurrence of drug withdrawal group were2 (0-6),16 (2-53),106 (14-254) and 156 (28-395) spot forming cell (SFC)/106 peripheral blood mononuclear cell (PBMC),respectively.The differences between healthy control group and immune-tolerance group,sustained antiviral treatment group or recurrence of drug withdrawal group were all statistically significant (Z=-3.585,-4.069 and-3.824,respectively;all P＜0.01).The IFN-γ level of HBcAg-specific CD8+ T cells in recurrence of drug withdrawal group was significantly higher than that in sustained antiviral therapy group (Z=-2.205,P=0.027),and that in sustained antiviral therapy group was significantly higher than that in immune-tolerance group (Z=-4.700,P＜ 0.01).The TNF-α levels secreted by HBcAg-specific CD8+ T cells in each group were 2 (0-5),16 (2-32),112 (15-283),and 195 (55-537) SFC/106PBMC,respectively.The differences between healthy control group and immune-tolerance group,sustained antiviral treament group or recurrence of drug withdrawal group were all statistically significant (Z=-3.619,-4.069 and-3.824,respectively;all P＜0.01).The TNF-α level secreted by HBcAg-specific CD8+T cells in recurrence of drug withdrawal group was significantly higher than that in sustained antiviral therapy group (Z=-2.449,P=0.014),and that in sustained antiviral therapy group was significantly higher than that in immune-tolerance group (Z=-4.350,P＜0.01).Conclusions The changes of frequency and immune function of HBcAg-specific CD8+T cells in CHB patients may be one of the reasons causing severe liver damage after irregular withdrawal of nucleoside analogues.

Objective To study the In vitro expansion of human CD8+ CD28- suppressor T cells and their immunological regulatory effect.Methods Human CD8 + CD28- suppressor T cells were expanded in vitro driven by the combination of cytokines and allogeneic antigen presenting cells (APCs).Flow cytometry was used to assess the development of CD28- subpopulation.Expanded CD8+ CD28- T cells were isolated by immunomagnetic microspheres and then added as third part modulators into mixed lymphocyte culture to assess their immunological regulatory characteristics.Results The combination of cytokines included IL-2,IL-7 and IL-15 and allogeneic APCs could increase the portion of CD8 + CD28- T cell subtype,and expansion fold of CD8+ CD28- T cell subtype was significantly increased as compared with others (P＜0.05).Expanded CD8+ CD28- T cells could suppress the proliferation of CD4+ T cells stimulated by allogeneic APCs.Moreover,this suppression had antigen specificity.Conclusion Human CD8 + CD28- suppressor T cells can be in vitro expanded in large amounts driven by the combination of cytokines and allogeneic APCs.Expanded CD8 + CD28- T cells in this study have antigen specific regulatory characteristics.

BACKGROUND: Many experiments have proved that heavy-load movement training causes the acute increase of free radicals. The increase of endogenous free radicals and caused cellular and subcellular lipid peroxidation strengthening injure the structure and function of tissue cells, thereby, decrease motor ability. Sleep deprivation also causes the increase of oxygen free radicals.OBJECTIVE: To observe the changes in malondialdehyde (MDA) and glutathione (GSH) levels as well as superoxide dismutase (SOD) activity in serum of acute exhaustive exercise rats following different time periods of sleep deprivation.DESIGN: A randomized controlled animal experiment.SETTING: Laboratory of Exercise Sciences & Sports Medicine, Physical College, Hunan Normal University.MATERIALS: Thirty healthy male SD rats of clean grade, weighing about (220±13)g, provided by Experimental Animal Center of Hunan Agricultural University, were involved in this study.METHODS: This experiment was carried out in the Laboratory of Exercise Sciences & Sports Medicine, Physical College,Hunan Normal University from April 2006 to May 2006. Thirty rats were randomized into 5 groups: blank control group,simple exercise group, sleep deprivation 24 hours group, sleep deprivation 48 hours group and sleep deprivation 72 hours group, with 6 rats in each group; Rats in the blank control group were allowed to sleep normally, but not do exercise; Rats in the simple exercise group were allowed to sleep normally and executed after acute exhaustive exercise; Rats in the sleep deprivation 24, 48 and 72 hours groups were deprived their sleep for 24, 48 and 72 hours,respectively, then they were executed after acute exhaustive exercise. Method of gentle handling was used in creating rat models of sleep deprivation; Rats in the simple exercise group and sleep deprivation groups were forced to do exercise according to the rat exercise model project established by Bedford: treadmill gradient 10°, speed 19.3 m/min, all the exercise rats were exhaustive (Exhaustion criteria: At the end of exercise, rats reached 1/3 of runway over 3 times;Various stimulations for expelling were invalid. After running, rats presented with breathlessness, expression lassitude,ventral decubitus, slow stimulus response, weaker escape response in being captured). After experiment, rats in each group were executed under the anesthetic state, and their blood was taken out, and centrifuged after natural clotting.Supernatant fluid was taken for detecting MDA and GSH levels as well as SOD activity.MAIN OUTCOME MEASURES: Changes in MDA and GSH levels, and SOD activity in rat serum.RESULTS: Thirty rats were involved in the final analysis. ① MDA level in the simple exercise group was higher than that in the blank control group (P＜ 0.01). MDA level in the sleep deprivation 24 hours, 48 hours and 72 hours was higher than that in the simple exercise group, respectively (P ＜ 0.01). MDA level in the sleep deprivation 72 hours was statistically higher than that in the other sleep deprivation groups, respectively (P ＜ 0.01). ②GSH level in the simple exercise group was lower than that in the blank control group (P ＜ 0.01). GSH level in the sleep deprivation 24 hours group was higher than that in the simple exercise group [(P ＜ 0.01). GSH level in the sleep deprivation 24 hours and 48 hours groups was lower than that in the simple exercise group, respectively (P ＜ 0.05, 0.01). There were statistical differences in GSH level among sleep deprivation groups (all P＜ 0.01). ③ SOD activity in the simple exercise group was lower than that in the blank control group (P ＜ 0.01); SOD activity in the sleep deprivation 24, 48 and 72 hours groups was lower than that in the simple exercise group, respectively (P ＜ 0.01); SOD activity in the sleep deprivation 48 and 72 hours groups was significantly lower than that in the blank control group, respectively (P ＜ 0.01); There were statistical differences in SOD activity among sleep deprivation groups (P ＜ 0.01).CONCLUSION: Sleep deprivation can cause serumal oxidative stress injury of rats; With elongation of time of sleep deprivation and exhaustive exercise, oxygen free radical production in serum of rats accumulates more and more, ability to get rid of oxygen free radical becomes weaker and weaker, and injury to body is more and more obvious.Serumal oxidative stress status of acute exhaustive exercise rats following sleep deprivation

Objective: To study left ventricular diastolic function (LVDF) in patients with coronary artery disease(CAD) by tissue Doppler imaging (TDI) and color Doppler echocardiography. Methods: The maximum velocity ( V max ) and time velocity integration(TVI) in early and late diastolic stage of mitral valve orifice of 80 patients with CAD and 79 normal subjects were measured by color Doppler echocardiography at apical 4 chamber view. Meanwhile, the V max and/or TVI of the myocardial motion spectrum of left ventricular lateral wall near the mitral valve annulet were measured by TDI. These data were compared and analysed by statistical method. Results: In the normal subjects, the older the subject, the lower the V max and TVI of early diastolic stage, the higher the V max and TVI of late diastolic stage. In CAD group, changes were not associated with ages and were affected by the degree of the myocardial pathology. The LVDF couldn't be exactly judged by color Doppler echocardiography when pseudonormalization blood flow spectrum appeared, but TDI technique was relatively sensitive. Any regional myocardial motion could be measured by TDI according to the examiner's design. Conclusion: LVDF by color Doppler echocardiography is simple and convenient. TDI technique can be used for determining overall LVDF and left ventricular systolic function. [

Objective Through investigating the effect of mesenchymal stem cells (MSCs) on the expression of yon Willebrand factor (vWF) and soluble thrumbomodulin (sTM) of vascular endothelial cells stimulated by serum of systemic lupus erythematosus (SLE) patients to discuss the repairing effect of MSCs on injured endothelium of SLE patients.Methods When human umbilical vein endothelial cell strain ECV-304 was co-cultivated with serum of SLE patients in vitro for twelve hours in order to induce endothelial cells injury,then MSCs were seeded for three days.Enzyme linked immunosorbent assay (ELISA) was used to detect the expression of vWF and sTM in the supernatant.Results When ECV-304 was stimulated by serum of SLE patients,the expression of vWF and sTM in the supernatant was significantly higher than that in the groups not stimulated by serum of SLE patients;When MSCs were seeded ,the expression of vWF in the Lupus serum stimulated group in which MSCs were seeded was significantly lower than that in the lupus serum stimulated group in which MSCs were not seeded,but the expression of sTM between the lupus serum stimulated group which MSCs were seeded and the lupus serum stimulated group in which MSCs were not seeded was not significantly deviatied.Conclusion Serum of lupus patients at active stage can cause injure vascular endothelial cells.MSCs can downregulate the expression of vWF of vascular endothelial cells.These suggest it may participate in the repairing of injured vascular endothelium of SLE patients.

Objective To observe the anti-atherosclerosis effects of Guizhi Fuling Capsule. Methods 50 healthy SD male rats were randomly divided into 2 groups: normal group (n=12) and AS model group(n=38). The model of experimental AS rats was established by feeding high cholesterol diet and intraperitoneal injecting VitD3. The normal control group was fed by ordinary diet. AS model group was further divided into 3 groups randomly: AS group(n=12), control group (n= 12), and therapy group(n= 12). The therapy group was given Guizhi Fuling Capsule by intragastric administration.The control group was given simvastatin and captopril by intragastric administration. 0.9% NaCl was given to the AS group instead of Guizhi Fuling Capsule. The rats were killed at the 16th weekend, and their aorta arch and breast aorta were observed.Blood fat and serum levels of Endothelin-1 (ET-1), Nitric oxide (NO), high-sensitivity C-reactive protein (hs-CRP) were detected respectively. Immunohistochemistry technology was used for observing the expression of intercellular adhesion molecule- 1 (ICAM- 1 ) in breast aorta. Results The values of ET- 1, NO, hs-CRP and the amount of ICAM- 1 were (142.61 ±25.67)ng/ml, (66.05 ± 8.63) μmol/L, (2.86±0.40)mg/L, (0.29±0.05) in the normal group; (182.38±22.96)ng/ml, (28.70±4.49)μmol/L, (5.60±0.49)mg/L, (0.58±0.06)in the AS group; (154.37±21.11)ng/ml, (45.88±11.36)μmol/L, (3.66±0.34)mg/L, (0.37±0.04)in the control group and (152.13±23.23)ng/ml, (57.67±8.96)μmol/L, (3.70±0.40)mg/L, (0.36±0.04) in the therapy group respectively. The levels of ET-1, NO, hs-CRP and the amount of ICAM-1 have significant difference between the AS group and the normal group(P=2.47E-4, 6.02E-19, 1.11E-12, 2.51E-18). There was also significant difference between the control group and the AS group(P=0.005, 1.59E-14, 1.87E-5, 3.87E-14). The level of NO in the therapy group was higher than that in the control group (P=0.002). The levels of ET-1, hs-CRP and the amount of ICAM-1 did not have significant difference between the therapy group and the control group(P=0.81, 0.81, 0.48) Conclusion Guizhi Fuling Capsule has effects of Anti-atherosclerosis on rats with AS.