Objective To investigate the effect of antisense oligodeoxynueleotide targeting sur-vivin (survivin ASODN) on hilar cholangioearcinoma cell line FRH-0201 depressing the expression of survivin. Methods Survivin ASODN was transfected into hilar eholangiocarcinoma cell line FRH-0201by liposome. Morphologieal changes were observed by inverted phase contrast microscope. RT-PCR and Western blot methods were performed to detect the expressions of survivin mRNA and protein re-spectively. The changes in cell apoptosis were detected by flow cytometry. Results The expression of survivin mRNA and protein was significantly decreased in survivin ASODN group than that in the con-trol group(mRNA: 0.51-t-0. 03 vs 0. 82-t-0.02,P%0. 05～protein: 1.82-t-0.16 vs 3. 08--t-_0. 27, P-Q 0.05). The morphologieal apoptotic changes were observed and the apoptosis rate was increased (11.50+1.49% vs 0.39-+-0.08～, P%0.05). Conclusion Survivin AS()DN can induce hilar cholan-gioeareinoma cell line FRH-0201 into apoptosis by decreasing the expression of survivin.

Objective To observe endoplagmic reticulum stress in bone tissue of fluomsis rats and further explore the pathogenesis of skeletal fluorosis.Methods 48 Wistar rats were divided into 4 groups according to their body mass.The control and low.calcium group were fed with normal diet(0.79%calcium)and low-calcium diet(0.79%calcium)respectively,and both drank tap water(sodium fluoride concentrations＜1 mg/L).High fluoride and low.calcium plus high-fluoride groups were fed with normal diet(0.79%calcium)and low-calcium diet (0.79%calcium)respectively,and both drank tap water containing sodium fluoride(sodium fluoride concentrations 221 mg/L).During experimental period,rats were measured body mass once a week with a stand diet and water available ad libiturn.The experimental period was 3 months.The biochemical techniques were used to test the indicators of oxidative stress and ALP in seFum of fluorosis rats.The total RNA was extracted from the one side of the femur,and the transcription level of Bip,Xbp1,CHOP and PDI were investigated by reverse transcription polymerase chain reaction(RT-PCR).Results The level of MDA in serum of low-calcium plus high-fluoride group wag higher than that of the control[(14.74±3.11)μmol/L vs(10.15±1.96)μmol/L,P＜0.05];the activity of GPx was ma~edly higher in hish-fluoride group compared with the control[(3.87±0.41)×103 U/L vs(2.85± 0.55)×103 U/L,P＜0.05];the level of uric acid in sel'um was significantly lower both in high-fluoride group and low-calcium plus high-fluoride group compared with the respective control and the low-calcium group[(73.95± 9.52)μmol/L vs(110.43±25.48)μmol/L,(54.32±22.09)μmol/L vs(101.71±17.01)μmol/L,P＜0.05]. The activity of ALP wag obviously higher in low-calcium plus high-fluoride group compared with the control [(24.77±4.57)×103U/L vs (12.91±3.97)×103U/L,P＜0.01)].The mRNA expression of Bip/GAPDH in bone tissue was markedly higher in bone of high-fluoride group and low-calcium plus high-fluoride group compared with the control(1.38±0.24,1.35±0.12 vs 1.14±0.06,P ＜ 0.05). The expression of Xbp1/GAPDH in bone tissue significantly increased in low-calcium plus high-fluoride groups compared with the control and the low-calcium group (1.48±0.20 vs 1.02±0.25,1.07±0.25,P ＜ 0.05 or ＜ 0.01);and CHOP/GAPDH in bone tissue significantly increased in low-calcium plus high-fluoride groups compared with the control(0.84±0.18 vs 0.52±0.07,P ＜ 0.05 ). Conclusions Accelerated osteogenetic action is seen in fluorosis rats,accompanied by oxidative stress and bone endoplasmic reticulum stress,which is likely involved in the pathogenesis of skeletal fluorosis.

Objective To study the effect of bromocriptine(BRC) on the activation of T lymphocyte stimulated by phytohemagglutinin(PHA).Methods After CD4+ T cell line Jurkat E6-1 cells were stimulated by PHA,prolactin(PRL) and BRC,respectively,the expression of linker for activation of T cells(LAT) and zeta-chain T cell receptor associated protein kinase 70 000(ZAP-70) mRNA of T lymphocytes were checked by RT-PCR.The expression of PRL mRNA of T lymphocytes was detected by Real time PCR.The expression of CD25(cluster of differentiation) as a marker of early activation on the surface of T lymphocytes was detected by flow cytometry,and the activation of nuclear factor-?B(NF-?B) was detected by luciferase reporter system.Results 1.BRC inhibited the expression of ZAP-70 as the common signal molecules both in the T lymphocyte activation pathway and PRL-prolactin-prolactin receptor(PRLR) signal transduction pathway,and decreased the expression of PRL mRNA produced by activation T lymphocytes.2.BRC enhanced the expression of LAT mRNA as another important signal molecular on the T lymphocytes and CD25 on the surface of the T lymphocytes.3.The activation of NF-?B of T lymphocytes was decreased.Conclusions BRC might inhibit the activation of T lymphocytes by inhibiting the expression of ZAP-70,the common signal molecules between T lymphocytes activation and PRL-PRL pathway,and PRL mRNA,the like-T lymphocyte growth factor.

Animals ; Cytokines ; blood ; Endotoxemia ; blood ; therapy ; Endotoxins ; blood ; toxicity ; Enzyme-Linked Immunosorbent Assay ; Hemofiltration ; Interleukin-1 ; blood ; Interleukin-10 ; blood ; Models, Animal ; Swine ; Time Factors ; Tumor Necrosis Factor-alpha ; analysis

Objective To investigate the expression of Caspase-9 and heat-shock protein-90 (HSP 90) in rats after focal cerebral ischemia-reperfusion injury.Methods The male SD rats (200-250 g) were divided into three groups by the random number table: normal group, sham group and cerebral ischemia-reperfusion (CIR) group.Each group was sorted into four subgroups including group 6 h, group 24 h, group 48 h and group 72 h according to the reperfusion time.Suture-occluded method was adopted to prepare focal cerebral ischemia-reperfusion(CIR) injury in rat model.Enzymelinked immunosorbent assay (ELISA) method was used to detect the variations of Caspase-9 and HSP-90 expression in rats.Results The changes in Caspase-9 and HSP 90 expression in the brain cells were observed by ELISA method.The expression of Caspase-9 and HSP-90 was weakly expressed in sham group, and was at peak in CIR group within 24 h-48 h, then began to decline at 72 h after the reperfusion time.The differences in the expression of caspase-9 and HSP-70 between sham group and normal group were not statistically significant.Conclusions Apoptotic cells gradually increase along with reperfusion time and reach the peak at 48 h after cerebral ischemia-reperfusion.In ischemia half dark stripe, the expression of Caspase-9 and HSP 90 is increased in neuronal cells after cerebral ischemia-reperfusion, and the positive cells number is at peak at 48 h after cerebral ischemiareperfusion.Apoptosis of neuronal cells after cerebral ischemia and reperfusion is a dynamic evolutionary process.The expression of Caspase-9 and HSP 90 in nerve cells plays an important role in regulating cell apoptosis.

Objective To observe the expression of vascular endothelial growth factor(VEGF) mRNA and protein in fluoride(F~-) treated fibroblast(FB) of mice in planar(2D) and FBs populated collagen lattice(3D) culture systems and to further explore the effects of VEGF on the osteogenic action of FB. Methods FB were divided into 0 (control group), 0.0001,0.0010,0.1000,1.0000,10.0000 and 20.0000 mg/L groups(F~-). The levels of VEGF mRNA and protein at 48 h were measured by using RT-PCR, ELISA and immunohistochemistry (IHC) methods. Results The expression of VEGF mRNA increased obviously in group of 0.1000 mg/L(1.08 ± 0.09) in 3D FB compared with the control group(0.93 ± 0.02, all P < 0.05). Fluoride increased the content of VEGF protein obviously in groups of 0.1000,1.0000,10.0000 mg/L(0.19 ± 0.02, 0.26 ± 0.01 and 0.32 ± 0.01 ), higher than that in 2D FB culture supematant in the control group(0.14 ± 0.01, all P < 0.05) ; and in groups of 0.1000, 1.0000 rag/L(0.59 ± 0.06 and 0.52 ± 0.03) it was higher than that in 3D FB culture supematant in the control group(0.37 ± 0.05, all P< 0.01 ). The IHC results showed that the VEGF positive staining cells increased significantly in group of 0.001 mg/L (0.45 ± 0.05) in 2D FB when it was compared with control group(0.36 ± 0.03, P< 0.05); and in groups of 0.0010, 0.1000, 1.0000 rag/L(0.62 ± 0.04,0.70 ± 0.06 and 0.65 ± 0.07) are it was higher than that in 3D FB control group (0.44 ± 0.04, P < 0.05 or < 0.01 ). Conclusions The higher expression of VEGF mRNA and protein in 2D and 3D FB induced by fluoride may play an important role in stimulating the osteogenesis ability in FB.

Calcaneus ; diagnostic imaging ; Female ; Humans ; Radiography ; Tuberculosis, Osteoarticular ; diagnosis ; diagnostic imaging ; Young Adult

10.3969/j.issn.2095-4344.2013.25.014

Objective To analyze the X-ray,CT and MRI findings of synovial tuberculosis,and to evaluate the role of MRI in diagnosing synovial tuberculosis.Methods Fourteen eases of synovial tuberculosis comfirmed by operation and pathology were retrospectively analyzed and summarized. All patients were examined by MRI and X-ray,and CT scans were performed in 3 cases. Results X-ray showed joint swelling(8 cases),articular space narrowing(7 cases),marginal joint erosions(4 cases), and periarticular osteoporosis(9 cases).The joint swelling was detected on CT in all 3 cases,and bony erosion and speckled sequestra were seen in 2 cases.MRI in all of patients showed joint swelling and synovial proliferation in different drgees,demonstrated as heterogeneously low signal on T_1 WI and slight high signal(7 cases)and obvious high signal(6 cases)on T_2WI,and diffuse synovial proliferation was demonstrated as massive and nodular signal in 8 cases.Joint effusion was present in 7 cases as low signal on T_1WI and high signal on T_2 WI.Osseous erosion lesions were seen in 7 cases,and intra-artieular cartilage thinned,partly or mostly disappeared in 11 cases.Periarticular bone marrow edema was found in 7 cases. Conclusion MRI was superior to X-ray and CT in the diagnosis and differential diagnosis of synovial tuberculosis.

Objective To identify the different causes of atrial fibrillation (AF) in Chinese by coronary artery angiography,and to compare the episode of stroke in different patients.Method A total of 782 adult patients diagnosed with AF were identified from Anzhen Hospital over a 8-year period.There were 273 patients with rheumatic valvular AF and 509 patients with nonvalvular AF.The results of electrocardiogram,echocardiography and coronary artery angiography were retrospectiwely analyzed to find out the proportion of causes and to compare the episode of stroke in patients with rheumatic valvular AF with those having nonvalvular AF.Results The patients with rheumatic valvular atrial were significantly younger than of coronary heart disease (52.40?5.03 years old vs 64.30?3.25 years old,P