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 .

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 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 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.

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 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.

Objective To investigate the possible pathological role of mitochondrial apoptosis pathways and its factors in fluorosis-induced apoptosis of human hepatocellular carcinoma cell strain (HepG2).Methods Under the stimulation of 1,3,6 and 9 mmol/L concentrations of NaF in vitro for 24 h (n =5),while normal control group was cultured under normal condition,the cytotoxicity was measured with MTT.The mitochondrial apoptosis inducing factor (AIF) was measured at both mRNA (n =5) and protein levels (n =6),respectively,by real-time PCR and Western blotting.The mitochondrial apoptosis related factors,such as B-cells lymphoma-2 (Bcl-2),Bcl-associated X protein (Bax),cytochrome C,caspase-9 and caspase-3 were measured at protein levels (n =6).Results After treated with 0,1,3,6 and 9 mmol/L NaF for 24 h,the cell absorbance of HepG2 cells was 0.307 ± 0.031,0.333 ± 0.028,0.230 ± 0.011,0.178 ± 0.001 and 0.152 ± 0.003,respectively,and the differences were statistically significant among groups (F =82.224,P ＜ 0.01).After treated with 3 mol/L NaF for 24 h,the mRNA level of AIF was [(153.14 ± 5.41)％] which was increased compared to the control group [(100.00 ± 4.70)％,t =-4.73,P ＜0.05].Under the same condition,the protein levels of AIF,Bcl-2,cytochrome C in cytoplasm,caspase-9 and caspase-3 were (152.16 ± 47.30)％,(171.90 ± 51.52)％,(458.00 ± 19.48)％,(527.17 ± 200.67)％ and (432.70 ±64.27)％,which were increased compared to those of the control groups [(100.00 ± 48.86)％,(100.00 ± 34.44)％,(100.00 ± 116.59)％,(100.00 ± 19.58)％ and (100.00 ± 137.16)％,t =-3.80,-3.96,-15.76,-4.64,-5.06,all P ＜ 0.05],while the protein levels of Bax and cytochrome C in mitochondrion were (24.66 ± 26.04)％,(72.99 ±45.34)％,which were decreased compared to those of the control groups [(100.00 ± 44.01)％,(100.00 ± 34.14)％,t =6.35,0.68,all P ＜ 0.05].Conclusion The mitochondrial apoptosis pathway and related factors may be involved in NaF-induced cell death in HepG2 cells.

Objective To report a new surgical approach (modified anconeus flap approach) in which the anconeus and part of triceps are reflected in the treatment of distal intercondylar humerus fracture (AO type C).Methods To design the modified anconeus flap approach,the anatomy of the distal tendon of the triceps brachii and the anconeus was studied using 15 cadaveric adult specimens (30 sides).The new surgical approach was compared with the triceps-reflecting anconeus pedicle (TRAP) approach in terms of the area of exposure at the distal humeral articular surface.Furthermore,the new surgical approach was applied in 16 patients who had been treated for intercondylar humerus fracture (AO type C) from May 2005 to May 2013.The clinical outcomes were evaluated using the Mayo elbow performance score (MEPS) and Broberg-Morrey scoring systems.Results The blood supply and innervation of the anconeus was interrupted minimally during incision along the radical edge of triceps tendon.The area of exposure at the distal humeral articular surface provided by the part triceps and anconeus reflecting approach and the medial humerus approach was 42.66％ ± 0.03％ at the elbow flexion from 0° to 50°.This was significantly smnaller than that provided by the TRAP approach (46.11％ ±0.03％) (P ＜0.05).Of the 16 patients,15 obtained complete follow-ups from 6 months to 4 years(average,16.5 months).The mean MEPS at the last follow-up was 90.5 points (range,from 82 to 93 points),with 10 excellent and 4 good cases and one poor case.The Broberg & Morrey score system showed 11 patients with no articular cartilage degeneration (level-0),3 patients with level-1 degeneration,and 2 patients with level-2 degeneration.Conclusion The modified anconeus flap approach proposed in the present study provides clear surgical vision and needs no olecranon osteotomy or olecranon dissection or ablation of the major triceps brachii tendon for intercondylar humerus fractures,thereby leading to early active motion and satisfactory clinical outcomes.

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 To investigate the influence of intravenous infusion by double-lumen peripherally inserted central catheter (PICC)on continuous dynamic central venous pressure(CVP)monitoring.Methods In the patient undergoing the double-lemon PICC,continuous dynamic CVP monitoring was performed by a lumen and the intravenous infusion was conducted by another lu-men.Whether the simultaneous venous infusion affecting the values of continuous dynamic CVP monitoring was observed.Results Intravenous infusion through double-lumen PICC had no influence on simultaneously continuous dynamic CVP monitoring (P>0.05),which could influence the values of CVP monitoring when simultaneously using the intravenous infusion pump(P<0.05). The speed of pumping fluid had no influence in a shout time(10 min),with significant difference(P>0.05).The CVP values could recovered to the status before infusion when continuously infusing after stopping using the infusion pump.Compared with before in-fusion,the CVP values after stopping intravenous infusion had no obvious change(P>0.05).Conclusion Intravenous infusion by double-lumen PICC has no influence on continuous dynamic CVP monitoring.But simultaneously adding infusion pump has certain influence on the CVP monitoring values.