Aim To study the effects of amitriptyline(Ami)on focal cerebral ischemia-reperfusion injury in rats.Methods An animal model of focal cerebral ischemia-reperfusion injury was induced by the middle cerebral artery occlusion(MCAO) by reversibly inserting a nylon thread method.The rats were decapitated after ischemia for 1 hour and reperfusion for 2 hours.The infarct volumes were determined using a 2,3,5-tri-phenyl tetrazolium chloride(TTC) staining and assessed by image analysis system.The neurologic deficit status were evaluated on 0~5 grade scale.The levels of dopamine(DA),norepinephrine(NE),serotonin(5-HT) and its metabolic product~hydroxyindole acetic acid(5-HIAA) in cortex and striatum were measured by fluoro-spectrophotometry.Results Ami treatment exhibited a remarkable reduction in infarct volume and neurologic deficit scores.The monoamines content of cortex and striatum had a significant increase compared with ischemia-reperfusion group.Conclusion Amitriptyline has protective effect on cerebral ischemia-reperfusion injury in rats.The mechanism might be related to reducing the release of NE,DA and 5-HT during cerebral ischemia-reperfusion,attenuating or inhibiting of the neurotoxic effects of monoamine neurotransmitters.

Objective To analyze the central effect of Qi-Yin Deficiency Syndrome (QYDS) scoring in clinical trials of diabetes.Methods Dispensation degree analysis and multiple linear regression analysis were adopted to compare the differences of central effects among the Qi-Yin deficiency syndrome scoring,fasting plasma glucose (FPG) level and 2-hour postprandial blood sugar (2 hPG)level before and after treatment in the suited databases from two phase Ⅲclinical trials of type two diabetes performed in 2004～2005.Results The variation coefficients of QYDS scoring treated with drug A and drug B varied from one tenth to half times of those of FPG and 2 hPG levels before and after treatment.And the influence of centers on QYDS is weaker than those on blood sugar levels (FPG &2 hPG) either referring to the numbers of significant centers or referring to the absolute values of standard regression coefficients in multiple linear regression equation.Conclusion There exists a central effect in QYDS scoring before and after treatment,and the central effect of QYDS scoring is weaker than that of blood sugar levels (FPG &2 hPG) in clinical trials of diabetes.

Objective To explore the semitivity of ovarian cancer cell line SKOV3 to paclitaxel,oroteasome inhibitors,bortezomib,and their combination.Methods The methyl thiazolyl tetrazolitim (MTT)assay was applied to examine the cell viability after treatment.The annexin V-propidium iodide apoptosis detection kit was used to determine the apoptosis rate of different groups.Western blot assay was used to evaluate the expression levels of phosphorylated protein kinase B(AKT)and glycogen synthase kinase-3 beta(GSK-3β).Results In MTT assay,the cell viability ratios of the combination group at serial time points from 12,24,36,48 and 72 hours Were(65.2±5.8)%,(58.3±14.4)%,(35.3±5.0)%,(19.2±1.5)%,and(11.4 ±2.5)%,which were significantly lower than those of the paclitaxel group (P<0.05).After arug treatments,apoptosis rates of paclitaxel group,bortezomib group and the combination group were (14.7±0.5)%,(15.1±0.8)%and(20.5±0.7)%respectively.The rate of the combination group was significantly higher than that of non-treated group and paclitaxel group(P<0.05).Western blot assay showed the changes in expression levels of phosphorylated AKT and GSK-3β,which were decreased significantly after paclitaxd and bortezomib combination treatment [(3.2±0.8)%,(19.3±0.4)%;P<0.05].Conclusions The lethal effect of paclitaxel on tumor cells could be increased significantly by its combination with proteasome inhibitors,bertezomib.The AKT/GSK-3β signaling pathway plays an important role in the molecular mechanism of the combination treatment.

Objective To explore the sensitivity and the molecular mechanism of cisplatinresistance ovarian cancer cell line C13 to proteasome inhibitors and the combination with cisplatin. Methods After different treatments, methyl thiazolyl tetrazolium (MTT) assay was applied to examine the cell viability, annexin-V/propidium iodide(PI) apoptosis detection kit was used to determine the apoptosis rate of different groups, western blot assay was introduced to evaluate the expression levels of Fas-associated death domain-like interleukin-1 beta converting enzyme inhibitory protein (cFLIPs), and the activity of caspase-8 was examined. Results MTT assay shown that the cell viability ratios of combination group at serial time points from 12, 24, 36, 48, 60, 72 hours were ( 56.0 ± 8.4 ) %, (44.7 ± 7.3 ) %, ( 33.7 ±11.2) %, (27.6 ± 8.0) %, (27. 6 ± 7.6) % and (28.1 ± 2.4) %, which were much lower than those of cisplatin group (P <0.05). After treated for 24 hours, apoptosis rates of cisplatin group, bortezomib group and combination group were ( 16.7 ± 1.7) %, (23.4 ± 2.1 ) % and (26.9 ± 1.6) %, respectively. The rate of combination group was much higher than that of non-treated group and that of cisplatin group or bortezomib group ( P < 0.05 ). Western blot assay showed the changes of expression levels of cFLIPs, which were downregulated seriously after cisplatin, bortezomib or combination treatment [ (43.2 ± 2.3 )% vs( 75.7 ± 3.0)%vs (67.9 ± 2.1 ) %, P < 0.05 ]. The caspase-8 activity of combination group was (5.6 ± 1.6) folds than that of non-treated group, which was higher than those of other two groups [ ( 2.3 ± 1.0) and (4.2 ± 0.9 ) folds,P < 0.05 ]. Conclusions The tumor cell lethal effect of cisplatin could be increase significantly by the combination application of proteasome inhibitors, bortezomib. And the cFLIPs/caspase-8 signaling pathway may be play an important role in the molecular mechanism of the combination treatment.

Multiple outcomes measured repeatedly for the same subject are common in longitudinal observation.If we use the approach by analyzing each outcome separately,it may lead to wrong conclusions due to the failure of accounting for joint evolution of different outcomes.To adequately capture the interdependence among multiple outcomes,we proposed a joint modeling for multivariate longitudinal data by constructing a linear mixed-effects model for each outcome and accommodating the relationship among multiple outcomes through correlation in random effects.Maximum likelihood method was adopted to estimate parameters in this model.The application of this method was demonstrated through the analysis of stroke data.

Although the combination of surgery, radiotherapy and chemotherapy is the main methods of cancer treatment, it still fails to solve certain tumors, especially metastatic tumors. With the in-depth study of tumorigenesis and development mechanism, and the exploration and clinical application of tumor immunotherapy, the survival period of patients with malignant tumors has been significantly prolonged. Tumor immunotherapy has become an effective anti-tumor method by activating the body′s own immune system to achieve tumor suppression. The combination of chemotherapy and immunotherapy has a significant effect and has become a feasible solution for cancer treatment. The rationally designed nanomedicines can effectively combine chemical drugs and immunological preparations, and have become an effective delivery carrier basis and treatment means for clinically targeting tumor tissues, synergistic immune mechanisms to kill tumor cells, and treating tumors. In this paper, the types of multifunctional nanomedicines used in chemotherapy and various immunotherapies in recent years and their advantages in cancer therapy were reviewed.

The biomimetic strategy of using the cell membrane-coated nanoparticles can retain the physical and chemical properties of the nanoparticles and show the biological characteristics of the source cell membrane, which can further enhance the role of the nanodrug in tumor treatment. A hybrid cell membrane is the fusion of two or more different types of cell membranes. A hybrid cell membrane can endow nanoparticles with multiple biofunctions derived from the source cells compared with a single cell membrane. Hybrid cell membranes provide a foundation to stimulate extensive research into multifunctional biomimetic nano-drug delivery system (NDDS), which is expected to broaden the application of biomimetic nanotechnology in drug delivery systems. In this review paper, the types of hybrid cell membrane used to construct nano-drug delivery systems, the preparation and characterization methods, and cancer treatment research progress in recent years were reviewed.

MicroRNAs (miRNAs) play critical roles in the development and progression in various cancers.Dysfunctional miR-9 expression remains ambiguous,and no consensus on the metastatic progression of ovarian cancer has been reached.In this study,results from the bioinformatics analysis show that the 3'-UTR of the E-cadherin mRNA was directly regulated by miR-9.Luciferase reporter assay results confirmed that miR-9 could directly target this 3'-UTR.miR-9 and E-cadherin expression in ovarian cancer tissue was quantified by qRTPCR.Migration and invasion were detected by wound healing and Transwell system assay in SKOV3 and A2780.qRT-PCR and Western blot were performed to detect the epithelial-mesenchymal transition-associated mRNA and proteins.Immunofluorescence technique was used to analyze the expression and subcellular localization of Ecadherin,N-cadherin,and vimentin.The results showed that miR-9 was frequently upregulated in metastatic serous ovarian cancer tissue compared with paired primary ones.Upregulation of miR-9 could downregulate the expression of E-cadherin but upregulate the expression of mesenchymal markers (N-cadherin and vimentin).Overexpression of miR-9 could promote the cell migration and invasion in ovarian cancer,and these processes could be effectively inhibited via miR-9 inhibitor.Thus,our study demonstrates that miR-9 may promote ovarian cancer metastasis via targeting E-cadherin and a novel potential therapeutic approach to control metastasis of ovarian cancer.

This article mainly introduces the functional clustering methods and demonstrates its performance by the real analysis of Chinese medical Zong Qi data.The functional clustering analysis hypothesizes that the discrete time series observations are dominated by a continuous function of time,which can be expressed by infinite basis functions.Functional clustering methods include raw data method,filtering method and adaptive method.When dealing with the sparse data clustering analysis,raw data method encounters the difficulty of matrix calculation due to the lack of data on some time grids.Filtering method suits for full time data,while when facing missing data,the fitting curve is inaccurate so that the clustering outcome cannot be explainable.Adaptive method can be applied flexibly to both full time and sparsely sampled data.In the real analysis section,the adaptive method is used to cluster the sparsely sampled Chinese medical Zong Qi time series data,where the elderly individuals are divided into three clusters,the ones with high level of Zong Qi,the ones with moderate level and those with low level.The adaptive method performs well on clustering individuals.

Objective : To investigate the improvement effect and possible mechanism of cornus officinalis glyco- sides on MRL / lpr lupus nephritis． Methods : Forty mice lupus-prone MRL / lpr mice were divided into model group， cornus officinalis glycosides low-dose，medium-dose，high-dose group (27．5，55，110 mg / kg) ，with 10 mice in each group，and 10 C57BL /6 mice were taken as the control group．Gastric administration was carried out 1 time / day for 4 weeks．24-hour urine was collected before drug intervention and 2 and 4 weeks after intervention to determine the content of urine protein．After the drug intervention，mouse serum and kidney tissues were taken and the levels of serum creatinine (Scr) ，urea nitrogen (BUN) and anti-double-stranded DNA (dsDNA) antibodies were detected by kits．The levels of interleukin-6 ( IL-6) ，tumor necrosis factor-α ( TNF-α) and interleukin-1 β ( IL-1 β) in serum were detected by ELISA．The changes in peripheral blood T lymphocyte subsets were detected by Flow cytometry ; The changes of renal histopathology were observed by HE staining ; The protein expression levels of toll-like receptor 4 (TLR4) ，myeloid differentiation factor-88 ( MyD88 ) ，nuclear transcription factor ( NF-κB) p65，and p-NF-κB p65 in kidney tissue were detected by Western blotting. Results : Compared with the control group，the changes of renal histopathology of the model group was severe，and the levels of 24-hour urine protein，Scr，BUN and anti-dsD- NA antibodies，IL-1 β, IL-6 and TNF-α significantly increased (P＜0. 01) ，the peripheral blood CD4 + T / CD8 + T lymphocyte ratio was significantly reduced (P ＜0. 01) ，and the protein expression levels of TLR4，MyD88 and p- NF-κB p65 in kidney tissue were significantly up-regulated (P＜0. 01) ; Compared with the model group，the chan- ges of renal histopathology of each dose group of cornus officinalis glycosides were significantly improved，the levels of 24-hour urine protein，Scr，BUN and anti-dsDNA antibodies，IL-1 β , IL-6 and TNF-α were significantly reduced (P＜0. 01) ，the peripheral blood CD4 + T / CD8 + T lymphocyte ratio significantly increased (P ＜0. 01 ) ，and the protein expression levels of TLR4，MyD88 and pNF-κB p65 in renal tissue were significantly down-regulated (P < 0. 01) ．Among them，the high-dose group had the most significant changes in each index．While there was no signifi- cant difference in the expression of NF-κB p65 protein in the groups． Conclusion Cornus officinalis glycosides have a certain effect on renal inflammatory response in mice lupus-prone MRL / lpr mice，and the mechanism may be related to the inhibition of TLR4 / NF-κB signaling pathway.