Objective To construct the eukaryotic vector that expressing hepatitis B virus (HBV) S and the single fragment of variety chain (ScFv) of monoclonal antiboy against cytotoxic Tlymphocyte-associated antigen-4 (CTLA-4) and to analyze the immunological activity of recombinant S-ScFv protein.Methods The oringially constructed pSect2/ScFv4F10 and pSect2/S were double enzyme digested by Sfi I and Hind Ⅲ,respectively.Then the HBV S gene was cloned into the pSect2/ScFv4F10 vector.The pSect2/ScFv4F10 and pSect2/S-ScFv4F10 were expressed in Chinese hamster ovary (CHO) cells,and the expressed proteins were verified through sodium dodecyl sulfatepolyacrylamide gelelectrophoresis(SDS-PAGE)andWesternblotting.Afterultrafiltration concentration and affinity chromatography,the biological affinity of the expressed ScFv4F10 and S-ScFv4F10 proteins were examined by competitive enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) technology.The comparison between groups was done by One-way ANOVA.ResultsThe eukaryotic expression vector of pSect2/S-ScFv4F10was successfully constructed,and relative molecular mass of the expressed protein of S-ScFv4 F10 was about 52 000 that analyzed by SDS-PAGE and Western blotting.With the fixed concentration of 4F10-mAb against CTLA-4,the A570 value of the mixed reaction with purified CTLA-4 antigen gradually increased with the decrease of ScFc fusion protein proportion; when the molar ratio of ScFv,S-ScFv4F10∶4F10=2∶1,the competitiveinhibitionratesagainst 4F10conjugatedantigenwere72.6％and64.5％,respectively.The affinity constants of association kinetics for CTLA-4 mAb,ScFv4F10 and S-ScFv4F10 with CTLA-4 antigen were 7.29 × 108 mol/L,9.52 × 106 mol/L and 2.04 × 106 mol/L,respectively,and the dissociation constants of KD were 1.40 × 10-9 mol/L,1.05 × 10-7 mol/L and 4.91 × 10-7 mol/L,respectively.ConclusionsThe eukaryotic expression vector of pSect2/S-ScFv4F10is successfully constructed,and the recombinant protein of S-ScFv4 F10 has a fairly high affinity with CTLA-4 antigen.

Objective To investigate early nasogastric nutrition on patients with acute decompensated chronic heart failure,and observe its influence on patient's disease outcome.Methods From January 2012 to December 2013,100 cases with acute decompensated chronic heart failure were used as the research subjects,were randomly divided into experimental and control groups with 50 cases each group.Patients of two groups adopt expansion of blood vessels,strong heart,diuresis,and routine anti-infection treatment;Based on this scheme,experimental group adopt early nasogastric nutrition.The cardiac function index,the change of nutritional status and clinical curative effect and the result of follow-up within 1 year after discharge of two groups were compared.Results Left ventricular end-diastolic dimension (LVEDD) and left ventricular end-systolic dimension (LVESD) in two groups after treatment were decreased,and the descender of experimental group was superior to that of the control group with statistically significant difference (P ＜ 0.05).After treatment,left ventricular ejection fractions (LVEF) of two groups were improved,and the rise of experimental group was superior to that of control group with statistically significant difference (P ＜0.05).In the two groups after treatment,the serum brain natriuretic peptide (BNP) levels were significantly decreased,and the descender of the experimental group was more than that of control group with statistically significant difference (P ＜ 0.05).After treatment,body mass index (BMI),the level of serum prealbumin (PA),transferrin (TRF),and the total number of lymphocytes of two groups were increased significantly (P ＜ 0.05);and BMI,the level of serum PA and TRF,and the total number of lymphocytes of the experimental group were significantly better than those of the control group (P ＜ 0.05).The total effective rate of the experimental group was significantly higher than that of the control group (P ＜ 0.05).The exacerbation rates,readmission and mortality rate of experimental group follow-up within 1 year after discharge were significantly lower than that of the control group (P ＜ 0.05).Conclusions The early nasogastric nutrition on the patients with acute decompensated chronic heart failure was better able to improve patients' nutrition state,more conducive to patients with cardiac function recovery,improve the clinical curative effect,and reduce the exacerbation rates,and readmission and mortality rate.

OBJECTIVETo evaluate the outcomes of T3a prostate cancer with unfavorable prognostic factors treated with permanent interstitial brachytherapy combined with external radiotherapy and hormone therapy.

METHODSFrom January 2003 to December 2008, 38 patients classified as T3a prostate cancer with unfavorable prognostic factors were treated with trimodality therapy (brachytherapy + external radiotherapy + hormone therapy). The prescription dose of brachytherapy and external radiotherapy were 110 Gy and 45 Gy, respectively. The duration of hormone therapy was 2-3 years. The endpoints of this study included biochemical failure-free survival (BFFS), distant metastasis-free survival (DMFS), cancer-specific survival (CSS), and overall survival (OS). Survival curves were calculated using the Kaplan-Meier method. The Log-rank test was used to identify the prognostic predictors for univariate analysis.

RESULTSThe median follow-up was 71 months. The serum pre-treatment prostate-specific antigen (PSA) level ranged from 10.0 to 99.8 ng/ml (mean 56.3 ng/ml), the Gleason score ranged from 5 to 9 (median 8), and the percentage of positive biopsy cores ranged from 10% to 100% (mean 65%). The 5-year BFFS, DMFS, CSS, and OS rates were 44%, 69%, 82%, and 76%, respectively. All biochemical failures occurred within 40 months. The percentage of positive biopsy cores was significantly correlated with BFFS, DMFS, and OS (all P=0.000), and the Gleason score with DMFS (P=0.000) and OS (P=0.001).

CONCLUSIONST3a prostate cancer with unfavorable prognostic factors presents not so optimistic outcome. Hormone therapy should be applied to prolong the biochemical progression-free or metastasis-free survival. The percentage of positive biopsy cores and the Gleason score are significant prognostic factors.

Androgen Antagonists ; therapeutic use ; Brachytherapy ; Combined Modality Therapy ; Gonadotropin-Releasing Hormone ; agonists ; Humans ; Male ; Neoplasm Grading ; Prognosis ; Prostatic Neoplasms ; mortality ; pathology ; therapy ; Treatment Outcome

Objective To investigate the CT angiography techniques of the pulmonary artery and aorta at a time.Methods Twenty-three patients who underwent the 64-slice spiral CT examinations of pulmonary and aortic angiography at a time were analyzed retrospec-tively (Group A).Also,13 cases whose pulmonary angiography showed normal (Group B)and 13 cases whose aortic angiography showed normal (Group C)enrolled in this study as control groups.The contrast media dose and the radiation dose were analyzed among three groups.The abnormal blood vessels were observed,and the CT values of the normal pulmonary artery and aorta and the number of the pulmonary 4,5 grade were compared among all groups.Results The contrast media dose in Group A was more than that in Group B or Group C,but less than the sum of that in Group B and Group C.The radiation dose in Group A was equivalent to that in Group C,which was obviously less than the sum of that in Group B and Group C.Six of 23 cases were accurately diagnosed as pul-monary embolism,1 case as pulmonary malformation,4 cases as aortic dissection whose true and false lumens clearly showed with contrast,5 cases as aortitis,and 10 cases as aortic atherosclerosis.For the cases whose angiography showed normal,there was no statistically significance of the number of the 4,5 grade of pulmonary artery and the CT values of pulmonary artery and the aorta compared with those of the control group (P >0.05 ).Conclusion The 64-slice spiral CT can be used to complete pulmonary and aortic angiography at a time,and the contrast media dose and the radiation dose can be reduced obviously.

Mitochondrial oxidative stress has been recognized as a preliminary and critical factor that aggravates the pathological cascade of Alzheimer's disease, which induces the production of β-amyloid protein, upregulates the expression of phosphorylated tau protein and triggers oxidative damage to lipids, proteins and mitochondrial deoxyribonucleic acid. Central neurons are more vulnerable to oxidative stress than non-neuronal cells due to their high oxygen demand, abundant unsaturated fatty acids and antioxidant enzymes deficiency. On this account, this review introduces the causes of mitochondrial oxidative stress, and analyzes the important role of mitochondrial oxidative stress in the pathogenesis of Alzheimer's disease. Meanwhile, the review focuses on the design and intervention strategies of drug delivery systems targeting mitochondrial oxidative stress in neurons, aiming to provide new ideas for the prevention and treatment of Alzheimer's disease.

Objective To detect the ascites CD64 index of patients with decompensated cirrhosis ,and explore the value in the di-agnosis of spontaneous bacterial peritonitis (SBP) .Methods Decompensated cirrhosis with ascites patients were divided into non-SBP group ,SBP suspected group and SBP diagnosed group .Two control groups were composed of patients with ascites culture pos-itive and non-SPB group .SBP suspected group were further grouped according to abdominal pain and the percentage of peripheral blood neutrophils .The CD64 index was detected by flow cytometry .Compared the difference of ascites CD64 index after treatment , and analyzed diagnosis performance of procalcitonin in human peripheral blood and ascites white blood cell count of SBP .Results The level of the ascites CD64 index was significantly higher in SBP diagnosed group (179 .39 ± 65 .56)and SBP suspected group (115 .49 ± 58 .42)than that of non-SBP group(26 .88 ± 26 .05)(P<0 .01) .The differences of the ascites CD64 index between SBP suspected associated with abdominal pain group and SBP suspected associated with painless group was not statistically significant (P>0 .05) .The level of the ascites CD64 index in SBP suspected with percentage of peripheral blood neutrophils elevated group was increased significantly than non-elevated group(P<0 .05) .CD64 index level in effective treatment group was significantly lower than the ineffective treatment group(P<0 .01) .The area of the ascites CD64 index under the curve was greater than procalcitonin in human peripheral blood and ascites WBC count ,and had a higher sensitivity and specificity .Conclusion The detection process of the ascites CD64 index is fast and with less influential factors ,which provides a new choice for the clinical diagnosis of SBP .

Arachidonic acid is an essential fatty acid in human nutrition and a biogenetic precursor of the biologically active prostaglandins and leukotrienes. ?~5 desaturase catalyzes the ?~5 dehydrogenation of di-homo-?-linolenic acid to form arachidonic acid in biosynthetic pathway of arachidonic acid. Using real time PCR technology,the transcriptional expression levels of gene encoding ?5 desaturase in three Mortierella alpina strains M10,M6 and M23,and in different growth phase of high arachidonic acid yielding strain M6,were determined. Results showed that there was a distinct corelationship between mRNA transcript level of ?~5 desaturase gene and biosynthesis of arachidonic acid. Results indicated that ?~5 desaturase plays an important role in arachidonic acid biosynthesis.

Adult ; Female ; Hepatitis B, Chronic ; complications ; drug therapy ; Humans ; Interferon-alpha ; adverse effects ; therapeutic use ; Nephrotic Syndrome ; etiology

Objective To investigate the induction and function of circulating CD4+ CD25+ regulatory T cells(CD4+CD25+ Treg cells)in patients with chronic hepatitis B,and their role in the progress of chronic HBV infection.Methods Twenty two patients with chronic hepatitis B(CHB),11 patients with acute hepatitis B(AHB)and 20 healthy blood donators were enrolled in the study.After stimulating with recombinant HBV antigen or anti-CD3 mAb,the fraction of CD4+ CD25+Treg cells in peripheral blood mononuclear cells(PBMC)from different subjects was analyzed using flow cytometry.CD4+CD25+ Treg cells and effector cells(PBMC-Treg)were sorted out and cultured alone or together in vitro,and the cell proliferation and cytokines production were measured. Serum HBV titers,HBV markers and liver function were also evaluated.Results The fraction of circulating CD4+CD25+ Treg cells induced in PBMC from CHB patients under the stimulation of recombinant HBsAg was much higher than that in AHB patients(F=3.47,P＜0.05)and normal controls(F=4.98,P＜0.05).With HBsAg stimulation,cell proliferation and cytokines production (IFN-γand IL-10)of CD4+ CD25+ Treg cells either from patients or from normal controls was very low when cultured alone,and there was no significant difference between two groups(P＞0.05).Cell proliferation and IFN-γproduction of effector cells from AHB patients were higher than those from CHB patients,however,when effector cells were cultured with CD4+Cd25+Treg cells at a ratio of 5:1,the cell proliferation and IFN-γ production were significantly inhibited (F=4.66,5.24,P＜0.05).Conclusion Circulating nificantly suppress the production and IFN-γ production of effector T Cells,which may be associated with the progress of chronic HBV infection.

BACKGROUND:Studies have demonstrated that tetrandrine has protection on acute spinal cord injury,but the specific mechanism remains poorly understood.OBJECTIVE:To study the protection of tetrandrine on rat acute spinal cord injury and to study its mechanism from apoptosis pathway.METHODS:A total of 100 rats were randomly divided into 4 groups.All rats were prepared for spinal cord injury models using modified Allen method except that in the sham-surgery group.Methylprednisolone and tetrandrine was injected into rats in the methylprednisolone and tetrandrine groups by tail intravenous injection prior to and at 24,48 hours after model preparation.The same volume of physiological saline was injected in the sham-surgery and model groups.Basso-BeatUe-Bresnahan(BBB score)was recorded at 8 hours,1,3,7 and 14 days after model preparation.The morphological changes of spinal cord injury sites were observed by hematoxylin-eosin staining and the expressions of bcl-2 and bax were determined by immunohistochemistry.RESULTS AND CONCLUSION:The BBB score of methylpradnisolone and tetrandrine groups were significantly higher than that model group at 7 and 14 days(P<0.05),but there were no significant difference between the methylprednisolone group and tetrandrine group(P>0.05).Hematoxylin-eosin staining showed that the spinal cord injured severely at 3-7 days,the injury degree in the methylpradnisolone group and tetrandrine group was slighter than that of the model group,with smaller bax expression and greater bcl-2 expression(P<0.01).The findings demonstrated that,tetrandrine is able to protect neurons from apoptosis and promote the nerve function recovery by inhibiting the expression of Bax and promoting the expression of Bcl-2.Its effect is not inferior to methylprednisolone.