Objective To study the effects of Xianlingqianggukoufuye (XLQGKFY) on postmenopausal osteoporosis in ovariectomized female rats. Method 60 female Sprague-dawley rats in 7-month were used, 50 of them were ovariectomized and randomly divided into 5 groups:ovariectomized (OVX), OVX+ Nylestriol, OVX+XLQGKFY (high dose, middle dose, low dose), the others were sham-operated group. Rats were treated with drugs starting at 3rd day after the operation for 90 days. Double in vivo fluorochrome labeling was administered to all rats. At the end-point of study, the blood was collected to detecte contents of ALP, StrACP in serum, the fourth lumar vertebra (LV4) and femur bone sections were cut and stained for bone histomorphometric analyses, biomechanical analyses and BMP analyses. Result XLQGKFY decreased greatly the StrACP content, increased bone stiffness, bone M-load, improved bone biomechanical property. Conclusion XLQGKFY has a good preventive effect on postmenopausal osteoporosis which provides for clinical use.

Objective To examine ultrastructural features of the arteriole responsible for intracerebral hemorrhage or the perforating branches artery around hematoma in patients with hypertensive intracerebral hemorrhage and explore the mechanism of hypertensive intracerebral hemorrhage. Methods Twelve hypertensive patients with CT proved intracerebral hemorrhage underwent operation. The small artery specimens were obtained through cortex fistula and their ultrastructures were observed under the electron microscope. Results Twelve specimens including 4 cases of duty arteriolae and 8 cases of perforating branch arteriolae were collected, Different degrees of degeneration were observed in three layers of the arteriola in all 12 specimens. Changes in endothelial cells included endothelial cell necrosis, collapse, or fallen of from endomembrane, accompanied by degeneration of internal elastic membrane, such as uneven thickness, absence of intermittent and medial smooth muscle cell necrosis. Myofilaments in the cytoplasm were condensed to form a high electron-dense cytoplasm. No micro-aneurysm was observed. Conclusions The pathological changes of cerebral small artery walls load-bearing layer in hypertensive patients include internal elastic layer rupture, smooth muscle layer of degeneration, decreased elasticity and increased fragility. Small artery walls may rupture, resulting in bleeding under the condition of rapid dynamic changes of blood flow.

Objective To elucidate the expression diversity of IL-2R subunits in T helper(Th) cells under different activation condition and clarify its relationship with the response to IL-2 induced proliferation.Methods In vitro cultured Th1 cells were activated by plate bound anti-mouse CD3 McAb.In both activated and inactivated Th1 cells the expression of different IL-2R subunit was evaluated by real-time PCR and fluorescence staining,3H incorporation assay was adopt to measure the proliferation of the cells in response to IL-2 treatment,IL-2 affinity was determined via I125 labeled IL-2.IL-2Rα siRNA transfection was applied to knockdown CD25 expression in activated Th1 cells and its effect was confirmed by Western blot,IL-2 induced proliferation in IL-2Rα siRNA transfected Th1 cells were subsequently detected.CD4 cells were isolated from na(i)ve BALB/c mice and were also stimulated by plate bound anti-CD3 McAb.Cells were harvested at different days posterior to the stimulation,CD25 content and IL-2 induced proliferation were determined by flow cytometry and 3H incorporation assay respectively.Results In comparison with inactivated cells,only the expression of CD25but not other IL-2R subunits was significantly increased in activated Th1 cells.In accordance with elevated CD25,IL-2 affinity was also increased in activated Th1 cells,however,which resulted in decreased cell proliferation in response to IL-2 treatment.In activated Th1 cells,when CD25 expression was differently knockdown by different-dosed IL-2Rα siRNA transfection,the highest IL-2 response was observed in the cells with partially CD25 depression.Forin vitro activated na(i)ve CD4 cells the elevated CD25 expression gradually decreased and the highest proliferation response was detected at day 8 post stimulation upon IL-2 treatment.Conclusion Although CD25 is necessary for IL-2 induced proliferation,however,the excessively expressed CD25 lead to lower proliferation response.Not fully activated Thl or CD4 cells,but partially activated cells with lightly increased CD25 content possessed highest proliferation rate in response to IL-2 treatment.

Objective To clarify whether the regulatory effect of all-trans-retinoic acid (atRA) on the antigen presentation function of dendritic cell(DC) is tightly associated with NF-κB signaling pathway.Also to clarify atRA mainly affected differentiated DC or influence the differentiation procedure from bone marrow cell to DC.Methods MOG35-55 immunized C57BL/6J mice received administration of atRA or not,splenic DC and CD4 cells isolated from immunized mice were subjected to in vitro cross-culture,treated with IL-12 and IL-23 respectively.Th1 and Th17 polarization of stimulated CD4 cells were determined by intracellular staining and FACS analysis,while the production of their corresponding cytokines,IFN-γ and IL-17,were measured by ELISA.Bone marrow cells were isolated from the femurs of the na(i)ve mice,treated with IL-4 and GM-CSF with or without synergistic RA treatment.MHC Ⅱ and CD11c molecules on the DC were assayed by immune staining and FACS analysis,their antigen presentation functions were decided by the proliferation and cytokine production of the Th1 effecter cells stimulated by antigen pulsed DC.To investigate the status of the NF-κB signaling pathway,the amount of phospho-Ser536 NF-κB p65 in the whole DC lysate and total NF-κB p65 in the nucleus were detected by Western blot.Finally,selective RA receptor antagonists were studied to figure out which receptor was majorly involved in the regulatory effect of RA on DC.Results Splenic DC from RA treated mice showed significantly decreased function of presenting antigen to stimulate CD4 cell polarization and cytokine production.Compared with untreated control,RA in vitro treated DC showed decreased expression of MHC Ⅱ and CD11c on its cell surface,which was accompanied with depressed function of stimulating Th1 cell proliferation and IFN-γ production.Further study revealed that RA mainly affect the differentiation procedure from BMC to DC,however,has no significant effect on differentiated DC as the aspects of MHC Ⅱ,CD11c expression,responder cell proliferation and cytokine production were evaluated.Decreased amount of phosphor-Ser536 NF-κB p65 in the whole cell lysate and total NF-κB p65 in the nucleus was investigated in RA treated DC,and decreased antigen presentation function of RA treated DC always came together with diminished activation of NF-κB signaling pathway.Finally it was demonstrated that RARβγ antagonist,but not RARα antagonist,could entirely block the RA effect on DC.Conclusion Retinoic acid inhibit the differentiation of DC as well as decrease its antigen presentation function,which might be resulted from the inactivation of NF-κB p65 signaling pathway and mediated by RARβγ.

Objective To explore the A2AR activation after traumatic brain injury mechanisms and the role of excessive tau protein phosphorylation.Methods With no specific mice experiment research of specific pathogens, position in the left parietal cortex in mice, by the method of controllable cortical against brain trauma model model, 15 min after injury in mice abdominal injection of A2AR specific inhibitors ZM241385 or use A2AR knockout mice, testing the brain neuron loss and tau protein phosphorylation level;Use specific agonists CGS21680 activate the original generation of nerve cells in the hippocampus and the A2AR human neuroblastoma cells, using immunocytochemistry and immunofluorescence test tua protein phosphorylation level of change, to observe axon transport function of mitochondria.Results Immunohistochemical results accumulation of optical density analysis showed that inhibition of A2AR activation can significantly reduce after cerebral trauma Ser404 tua protein loci phosphorylation levels, reduce excessive tua protein phosphorylation with nerve pathological change;A2AR activation after tua phosphorylation of proteins at a Ser404 site level increased significantly, nerve axons per unit length processes in the mitochondria number decreased significantly, resulting in axoplasmic transport dysfunction;To activate the original generation of nerve cells in the hippocampus and after the A2AR human neuroblastoma cells, tua protein phosphorylation Ser404 locus levels increased significantly.Conclusion A2AR activation after cerebral trauma has obvious influence on tua protein phosphorylation levels, may be a function by influencing the axoplasmic transport, eventually forming cognitive dysfunction.

Objective To investigate the prevalence of toxoplasma gondii (Tox),rubella virus (RV),cytomegalovirus (CMV) and herpes simplex virus (HSV) infections (TORCH infections) among childbearing-age population in Henan province.Methods Enzyme-linked immunosorbent assay (ELISA) was applied to detect plasma TORCH IgM and IgG among 3084 childbearing-age men and women from theFirst Affiliated Hospital of Zhengzhou University during July and September,2011.The positive rates of anti-TORCH antibodies were compared among the various age and gender groups by x2 test.Results The total positive rate of anti-TORCH IgM was 5.5％ (170/3084),in which the positive rate of anti-RV IgM was the highest (2.9％),followed by anti-HSV IgM (1.0％).Within positive rate of anti-TORCH IgG,anti-HSV IgG was the highest (90.4％),followed by anti-CMV IgG (89.7％),RV IgG (48.1％) and Tox IgG (0.7％).The positive rate of anti-TORCH IgM was the lowest in individuals aged ＞ 30-40 year old.With the age increasing,the positive rates of anti-Tox IgG,anti-CMV IgG and anti-HSV IgG increased,but the positive rate of anti-RV IgG decreased.Women had higher positive rates of anti-CMV IgG and antiHSV IgG than men (x2 =83.470 and 7.026,P ＜ 0.O1).Conclusions Current infection of TORCH exists in childbearing-age population of Henan province,and the positive rate of anti-RV IgG is low.It is recommended to screen for TORCH infection in childbearing-age men and women.

The mRNA expressions of chemerin and its receptor CMKLR1 in adipose tissue of OLETF rats and LETO rats were detected by real-time PCR. The results showed that the two genes mRNA expressions in subcutaneous and visceral adipose tissue in OLETF group were significantly higher than those in LETO group (P< 0.05 or P<0.01) and expressions of them in visceral adipose tissue were higher than those in subcutaneous adipose tissue(P<0. 05 or P<0. 01), suggesting that chemerin and CMKLR1 may be involved in the pathogenesis of obesity, insulin resistance, and type 2 diabetes mellitus.

Objective To investigate whether toll like receptor ( TLR) signaling pathways can in-crease the expression of IL-17 R in neuralglial cells , and if they can whether the increased IL-17 R is func-tional.Methods Experimental autoimmune encephalomyelitis (EAE) was induced in B6 mice by immuni-zation with an emulsion of myelin oligodendrocyte glycoprotein 35-55 ( MOG35-55 ) in complete Freund's adju-vant (CFA).The expression of Il17ra and Il17rc in the brains and spinal cords of mice with EAE were de-tected by real-time PCR.Luxol fast blue ( LFB) staining was performed to the spinal cord sections to detect tissue demyelination.Immunohistological staining against IL-17RA and CD3 were undertook to visualize IL-17RA+and CD3 + cells.Same approaches were also applied to immunized Rag1 -/- mice to figure out whether T cells infiltration is necessary for increasing IL-17RA expression in the central nervous system ( CNS) .Then B6 mice were immunized with incomplete Freund′s adjuvant ( IFA) plus different TLRs ago-nists to measure the expression of Il17ra in the brains and spinal cords by qPCR .The purified astrocytes , microglia and oligodendrocytes isolated from neonatal mice brains were cultured in vitro for two weeks , and then treated with different TLRs agonists .The expression of Il17ra at mRNA and protein levels in the cells were determined by qPCR and Western blot respectively .The astrocytes were treated with IL-17A and LPS individually or in the combination to detect the level of CCL 2, CXCL8 and IP-10 in the supernatant by ELISA.Results B6 mice with induced EAE showed significantly increased Il17ra expressions in the brain and spinal cord , which was also detected in immunized Rag1 -/-mice.Although no spinal cord demyeliza-tion and CD3 cells infiltration were detected in Rag1 -/-mice, significantly increased number of IL-17RA positive cells could still be visualized .In vivo TLRs agonist participated immunization and in vitro treatment of purified neuroglial cells demonstrated that TLRs agonists could directly evoke IL -17RA expression in the CNS or cultured astrocytes , microglia and oligodendrocytes with high efficiency .Both IL-17 A and LPS could stimulate astrocytes to secrete CCL2, CXCL8 and IP-10, however, a combined use of IL-17A and LPS fur-ther augmented the production of these chemokines to a large extend .Conclusion Taken together , we con-cluded that TLRs agonists could directly stimulate neuroglial cells to express IL -17RA which functionally re-spond to IL-17A by secreting chemokines .

Objective:This study aims to investigate the correlation between the number of resected lymph nodes (LNs) and the prognosis of patients with node-negative non-small cell lung cancer (NSCLC). Methods:A retrospective review of 305 patients with NSCLC, who received curative resection between January 2004 and December 2009, was conducted. All patients were proved without lymph node involvement histopathologically. The prognostic impact of the number of negative LNs and the clinicopathologic factors were analyzed. Results:The overall median survival time and the 1-, 3-, and 5-year overall survival rates were 60.0 months, 76.1%, 59.3%, and 47.1%, respectively. Survival analysis confirmed that the number of negative LNs, T staging and the stations of the mediasti-nal lymph nodes dissected were showed to be independent prognostic factors. Patients with a high number of negative LNs had better overall survival than patients with a low number of negative LNs (P<0.05). The patients with dissected LNs counts of more than 11 for cases with pTl~2 tumor (P<0.05) and more than 16 for cases with pT3 tumor (P<0.05) had better long-term survival outcomes. The post-operative complication rate was 12.1%, which was not significantly correlated with the number of dissected lymph nodes (P>0.05). Conclusion:The number of negative lymph nodes is an independent prognostic predicting factor for node-negative NSCLC. Sufficient dissection of LNs is recommended to improve the survival of the patients with node-negative NSCLC.

Our earlier research has shown that mono-substituted N-phenyl-2, 2-dichloroacetamide exhibited much higher anti-cancer activity than the lead compound sodium dichloroacetate (DCA). In this paper, a variety of multi-substituted N-phenyl-2, 2-dichloroacetamides were synthesized and biologically evaluated. The results showed that 3, 5-disubstituted N-phenyl-2, 2-dichloroacetamide analogues had satisfactory potency. Among them, N-(3, 5-diiodophenyl)-2, 2-dichloroacetamide had an IC50 of 2.84 micromol x L(-1) against non-small cell lung cancer cell line A549 and could induce cancer cell apoptosis.