Administration, Oral ; Anti-Bacterial Agents ; administration & dosage ; Cytomegalovirus Infections ; drug therapy ; immunology ; virology ; Erythromycin ; administration & dosage ; Female ; Humans ; Infant ; Male ; T-Lymphocyte Subsets ; immunology ; Virus Replication

Objective To research the significanec of genes bel-2 and bax in chondrocyte apoptosis in experimental osteoarthritis in rabbits.Method Twelve New Zealand rabbits divided into two groups at random:model group and control group.Model group with osteoarthritis was duplicated by immobilizing with gypsum at extension position in right hind limb of rabbits.Rabbits of model group were executed after 6 weeks and chondrocytes were taken.Positive expression rate of bcl-2 and bax mRNA was measured by immunohisto- chemistry and Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL).Results Positive expression rate of mRNA in bel-2 and bax of model group was obviously more than control group(P＜0.05).According to immunohistochemistry,and grey grade of positive expression of protein in bcl-2 and bax of chondrocytes in model group was lower and positive expression rate was gigher compared with control group (P＜0.05,P＜0.01).bcl-2 and bax of model group were lower than control group(P＜0.05).Conclusion Genes bcl-2 and bax participate and thus accelerate chondrocyte apoptosis of osteoarthritis together.

Objective To investigate the phosphorylation of tau protein and the effect of atorvastatin on tau protein phosphorylation in hypercholesteremic mouse hippocampus. Methods Male Kunming mice were randomly divided into normal control group, hypercholesteremia group, and low-dose atorvastatin treatment group (8mg·kg-1·d-1), middle-dose group (15mg·kg-1·d-1),and high-dose group (20mg·kg-1·d-1) with five mice in each group. The hypercholesteremia mouse model was induced by cholesterol-enriched diets. The expression level of tau protein phosphorylation in mouse hippocampus was detected by Western blot and immunohistochemistry methods. The activities of mitogen-activated protein kinase (MAPK) and cyelin dependent kinase 5 (Cdk-5) were measured by liquid scintillation counting of the hippocampus incorporated radioactivity and immunoprecipetation activity assay respectively. Results In hypercholesteremic group, the expression level of tau protein phosphorylation in mouse hippocampus was significantly increased(F=14.761,P＜0.01)as compared with control group, and so were MAPK activity and Cdk-5 activity(all P＜0.01). Atorvastatin treatment group showed the decreased expression level of tau protein phosphorylation at ser396/404 site in low-dose group (F=6.349,P＜0.05),middle-dose group (F=10.283,P＜0.01) and high-dose group (F=10.511,P＜0.01) as compared with hypercholesteremia mouse group. The activities of MAPK and Cdk-5 were decreased along with the increasing atorvastatin doses. Conclusions Hypercholesteremia induces tau protein hyperphosphorylation in mouse hippocampus. Abnormal cholesterol metabolism may play an important role in the pathology process of neurodegeneration in brain. Atorvastatin might inhibit tau protein hyperphosphorylation by inhibiting the activations of MAPK and Cdk-5 in brain, atorvastatin may have the protect effect for tau protein.

Objective To evaluate the efficacy and safety of a levonorgestrel-releasing intrauterine system(LNG-IUS)for the treatment of dysmenorrhea associated with adenomyosis.Methods We recruited 48 women with moderate or severe dysmenorrhea associated with adenomyosis.All women were inserted of LNG-IUS into their uterine cavity from days 5-7 of their periods and maintained for 12 months.We compared the visual analogue scale(VAS)scores and verbal rating scale(VRS)scores of their dysmenorrhea and dyspareunia at baseline and 12 monthes follow-up.Results Forty-four women completed the study. There were significant differences between mean VAS and VRS scores changes of dysmenorrhea and dyspareunia at baseline and 12 monthes follow-up,those of dysmenorrhea dropping from 75?13 to 11?11 and 2.3?0.4 to 0.4?0.3,those of dyspareunia dropping from 54?19 to 4?4 and from 1.6?0.8 to 0.2?0.2 respectively.Overall 29 women(66%)were very satisfied or satisfied with the one-year treatment. Conclusion Insertion of LNG-IUS alleviates moderate or severe dysmenorrhea associated with adenomyosis remarkably.

Objective: To explore independent risk factors of coronary artery calcification (CAC) and analyze correlation among risk factors of CAC and serum osteopontin (OPN) level. Methods: According to results of 64-slice spiral computed tomography (MSCT) coronary angiography, a total of 65 patients were continuously enrolled and divided into CAC group (n=37) and non-CAC control group (n=28). Enzyme linked immunosorbent assay (ELISA) was used to measure serum level of OPN. Single factor and multiple factor Logistic regression analysis’s were used to analyze risk factors of CAC. Spearman’s straight line analysis was used to analyze correlation between risk factors of CAC and serum OPN. Results: 1、 The age, hypertension, diabetes, poor eating habits，lack of exercise, overweight, etc., which were independent risk factors of CAC （OR=3.47～12.96, P=0.018～0.003）by single factor Logistic regression analysis, were inducted to multiple factor Logistic regression analysis, its result showed that age, overweight, poor sleep quality, poor eating habits were independent risk factors of CAC, OR=35.31～5.17, P＜0.01～0.05; 2、Serum level of OPN in CAC group was significant higher than that of non-CAC control group ［(39.919±11.879) μg /L vs. (24.000±6.000) μg /L，P<0.01］; 3、The Spearman straight line correlation analysis indicated that serum level of OPN was correlated with risk factors of CAC : positively correlated with LDL-C, overweight, age, TC（r=0.487～0.286,P＜0.001～＜0.05）, and positively correlated with poor sleep quality, diabetes, poor eating habits, lack of exercise（r=4.10～2.24， P<0.01～0.05）; negatively correlated with HDL-C（r=-0.250,P＜0.05）. Conclusion: Correlation analysis indicates that age, overweight, poor sleep quality, poor eating habits etc. are independent risk factors of CAC；Serum OPN level is correlated with LDL-C, overweight，age, diabetes, lack of exercise etc., so these indicate that must decrease OPN level and risk factors of CAC to relieve CAC and slow down its development.

A total of 59 untreated asthmatics and 47 healthy control subjects were recruited from Qianfoshan Hospital of Shandong Province from May 2011 to April 2012.Compared with healthy control subjects,the levels of IL-25 in induced sputum and eosinophils,IgE,interleukin-4 (IL-4) and interleukin25 (IL-25) in serum samples of asthmatics were significantly higher while the level of interferon-gamma (IFN-γ) were much lower.However,after inhaled glucocorticoid treatment,the levels of eosinophils,IL-4 and IL-25 decreased and IFN-γ significantly increased,while the level of IgE showed no significant changes.It was also found the expression of IL-25 was markedly positively correlated with the levels of eosinophils and IL-4 in serum and markedly negatively correlated with the levels of IFN-γ and had no relatio.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent. Epigallocatechin-3-gallate (EGCG) is a polyphenolic constituent of green tea. In this study, inhibitory effect of combined use of EGCG and TRAIL on human melanoma A375 cells was examined and the possible mechanism investigated. The cells were divided into 4 groups: control group, EGCG group (EGCG: 10, 20 mug/mL), TRAIL group (TRAIL: 25 ng/mL) and EGCG+TRAIL group (combined group). The growth inhibition was measured in the A375 cells treated with different concentrations of TRAIL ((25, 50, 75, 100, 125, 150 ng/mL) by MTT assay. The apoptosis was assessed by flow cytometry. The expressions of DR4 and DR5 were detected by flow cytometry and western blotting. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that TRAIL could dose-dependently inhibit the growth of A375 cells and the IC(50) of TRAIL was 150 ng/mL. The apoptosis rate was 11.8% in the TRAIL group, 5%-7% in the EGCG group and 48.9%-59.1% in the combined group. Significant difference was found in the apoptosis rate between the combined group and the EGCG or TRAIL group (P<0.05 for each). The expression of DR4 instead of DR5 was significantly increased in the EGCG group. The activity of caspase-3 rather than caspase-8 was substantially enhanced in the EGCG group. These results suggest that EGCG is useful for the TRAIL-based treatment for melanoma.

Objective To investigate new type cytokine induced killer cells expansion using advanced breast cancer′s peripheral blood .Methods peripheral blood mononuclear cells were isolated from 8 advanced breast cancer volunteers and co -cultured with Cytokine induced killer cells .These cells were placed in plastic flasks containing CIK-MediumTM supplemented with 10% auto-plasma in the presence of IL -2 ( 1 000 IU/mL) .The cultures were fed with CIK-MediumTM supplemented with IL -2 following the proliferation capacity . Cell proliferation was measured by cell counting during the cultivation .Fourteen days after cultivation ,cell mark-ers CD3/CD16/CD56 were examined by flow cytometry .51Cr and MTT assays were employed in cytotoxicity as-says.Cytokines were assayed by ELISA method .Results CD16+,CD16+CD56+,CD56+CIK cells were 5.8~11.6%in 2 ×107 fresh PBMCs and 95.2~97.6%in co-cultured cells after 18 days cultivation .The in vitro ex-pansion rate of new type cytokine induced killer cells was up to more than 8.2 ×108 in total,the cytotoxicity are ef-fective killing cells against MCF 7 and BT20 breast cancer cell lines .New type cytokine induced killer cells expand-ed from all PBMCs and secreted cytokines IFN -and TNF-.Conclusion The present culture could be useful to clarify the mechanisms of CIK cells expansion in vitro and feasible for breast cancer immmuno cell therapy .

Objective To investigate the correlation of plasma interleukin ( IL)-17 level and IL-17 receptor (IL-17R) expression in the thymus of patients with myasthenia gravis (MG).Methods The blood samples of 63 patients (38 with glucocorticoid treatment, 25 with thymus removal) who admitted to Henan Provincial People′s Hospital between 2010 and 2014 were collected at three different stages: pre-treatment, 1 week post-treatment and 1 month post-treatment.The blood samples of 42 healthy controls were also collected.Enzyme linked immunosorbent assay was used to evaluate the levels of IL-17 in plasma.Twenty-five thymus tissues from MG patients and another 12 thymus tissues from patients with congenital heart disease who had surgery therapy were also collected.Reverse transcription polymerase chain reaction was used to evaluate the mRNA levels of IL-17R.The possible correlation between the expression of IL-17 and IL-17R with MG was analyzed.Results Before treatment, the levels of IL-17 in the plasma were much higher in all the MG patients ( both ocular and generalized) when compared to the healthy controls ( controls (3.2 ±0.7) pg/ml, MG patients (8.5 ±1.7) pg/ml, t =2.450, P <0.01; generalized type patients (9.7 ±1.4) pg/ml, t =2.532, P <0.01).In the patients with glucocorticoid treatment, IL-17 levels began to reduce after 1 week treatment and a statistically significant difference was found when compared to the pre-treatment samples (pre-treatment (8.3 ±1.2) pg/ml, 1 week after treatment (6.3 ±0.7) pg/ml, t=2.052, P<0.05) and healthy controls (t =1.933, P<0.05).One month after the glucocorticoid treatment, the levels of IL-17 decreased to the normal level (1 month after treatment (3.9 ±0.6) pg/ml, t=2.630, P <0.01, compared to the pre-treatment; t =1.395, P >0.05, compared to the healthy controls).In the surgery therapy cases, the IL-17 levels were also reduced after the thymus removal ( pre-surgery (8.8 ±1.4) pg/ml, 1 week after surgery (5.3 ±0.7) pg/ml, t=1.950, P<0.05;1 month after surgery (3.0 ±0.4) pg/ml, t=2.683, P<0.01).In the thymus tissues of the MG patients, the mRNA levels of IL-17R were much higher than that of the controls ( relative level 2.31 folds, t =2.682, P <0.01).Meanwhile, a positive correlation was found between the plasma IL-17 levels and the relative IL-17R levels in thymus tissues ( r =0.945 4, P <0.01 ).Furthermore, IL-17 was positively correlated with quantitative myasthenia gravis scores (QMGS) either pre-treatment (r =0.798 1, P <0.01) or post-treatment (r=0.906 5, P<0.01).And IL-17R was positively correlated with QMGS pre-treatment (r=0.775 5, P<0.01).Conclusions IL-17 is increased in the plasma of MG patients (both ocular and generalized) , and is decreased upon the glucocorticoid treatment or surgery therapy, suggesting that it can be used as a parameter to determine the therapeutic effects.IL-17R is increased in the thymus tissues of MG patients, suggesting that it can potentially be used as a pathological diagnosis parameter.

BACKGROUNDAlzheimer's disease (AD) is a neurodegenerative disorder and the leading cause of dementia in the elderly. The two hallmark lesions in AD brain are deposition of amyloid plaques and neurofibrillary tangles (NFTs). Hypercholesteremia is one of the risk factors of AD. But its role in the pathogenesis of AD is largely unknown. The aim of this study was to investigate the relationship between hypercholesteremia and tau phosphorylation or beta-amyloid (Abeta), and evaluate the effect of atorvastatin on the level of tau phosphorylation and Abeta in the brains of rats fed with high cholesterol diet.

METHODSSprague-Dawley (SD) rats were randomly divided into normal diet control group, high cholesterol diet group, and high cholesterol diet plus atorvastatin (Lipitor, 15 mg x kg(-1) x d(-1)) treated group. Blood from caudal vein was collected to measure total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL) and high-density lipoprotein (HDL) at the end of the 3rd and the 6th months by an enzymatic method. The animals were sacrificed 6 months later and brains were removed. All left brain hemispheres were fixed for immunohistochemistry. Hippocampus and cerebral cortex were separated from right hemispheres and homogenized separately. Tau phosphorylation and Abeta in the brain tissue were determined by Western blotting (using antibodies PHF-1 and Tau-1) and anti-Abeta40/anti-Abeta42, respectively.

RESULTSWe found that high cholesterol diet led to hypercholesteremia of rats as well as hyperphosphorylation of tau and increased Abeta level in the brains. Treatment of the high cholesterol diet fed rats with atorvastatin prevented the changes of both tau phosphorylation and Abeta level induced by high cholesterol diet.

CONCLUSIONSHypercholesteremia could induce tau hyperphosphorylation and Abeta production in rat brain. Atorvastatin could inhibit tau hyperphosphorylation and decrease Abeta generation. It may play a protective role in the patho-process of hypercholesteremia-induced neurodegeneration in the brain.

Administration, Oral ; Amyloid beta-Peptides ; metabolism ; Animals ; Antibodies, Monoclonal ; Atorvastatin Calcium ; Blotting, Western ; Brain ; drug effects ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Heptanoic Acids ; administration & dosage ; pharmacology ; therapeutic use ; Immunohistochemistry ; Male ; Phosphorylation ; drug effects ; Pyrroles ; administration & dosage ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; tau Proteins ; metabolism