Objective To investigate the regulation on cell cycle related factor such as Cyclins and P21waf/cip1 by inhibiting histone deacetylase(HDAC)with valproic acid sodium(VPA).Methods HepG2 hep-tocellular carcinoma cells.BGC-823 gastric carcinoma cells and MCF-7 breast cancer cells were cultured with O.75-4.00 mmoL/L VPA for 48 h in vivo.and the cell cycle was analyzed by flow cytometry with PI assay.The protein and mRNA expression of Cyclin A,Cyclin D1,Cyclin E and P21waf/cip1 were analyzed by indirect immu-nofluorescence technique and RT-PCR.respectively.Results Compared with control groups,VPA at concen-trations 0.75-4.00 mmol/L exerted a significant inhibiting effect on G1 phase of HepG2,BGC-823 and MCF-7 cells(P<0.001).and the effect was dose dependent.Cyclin A was down-regulated both at mRNA and protein level in HepG2 and BGC-823 cells(P<0.001),but no difference in MCF-7 cells(P>0.05).Cyclin D1 was down-regulated both at mRNA and protein level(P<0.001)and P2lwaf/cip1 was up-regulated both at the mRNA and protein level in the three cell lines(P<0.001);Conversely,protein and mRNA expression of Cyclin E were unchanged upon treatment with VPA(P>0.05).Condusion Acetylization of histone intervened with VPA can regulate Cyclin D1 and P21waf/cip1 expressions obviously.To the expression of Cyclin A,it shows some difference according to the histogenesis and phenotypes of different carcinoma types.But there is not any obvious function on Cyclin E.Down-regulating Cychn D1 and up-regulating P21waf/cip1 may be the common target path-way in the inhibition of cell cycle G1 phase exerted by VPA.

Objective:To study the varieties and contents of related substances in cefalotin sodium by HPLC and MS. Methods:The quantitative determination of impurity spectra was carried out using self-contrast method, and UPLC-MS was used to preliminarily i-dentify the variety of impurities. Results:Through the study on the ascription and content of the related substances, there were 6 sub-stances existing in cefalotin sodium, and through the structure analysis, the 6 substances showed antibiotic action and toxicity. Conclu-sion:The method is simple, stable and repeatable, which can be used for the determination of cefalothin sodium and related sub-stances.

Purpose To retrospectively analyze and summarize the image features of rare ovarian clear cell carcinoma (OCCC), and to provide basis for clinical diagnosis. Materials and Methods CT and MRI features of 30 cases of OCCC confirmed by pathology were retrospectively analyzed. Sixteen of all the patients underwent pre- and post-contrast CT scan. And 14 patients underwent pre- and post-contrast MRI scans. Results ① CT features: unilateral mass was revealed in 15 cases and bilateral mass was in 1 case. The maximum diameter of the mass ranged from 8 to 23.7 cm, mean (12.86±3.96) cm. One mass was irregular, 4 masses revealed incomplete capsule, and 4 masses had septa in the mass. CT value of cystic part of the mass was 20-30 HU, and which of solid part was 28-53 HU. On post-contrast CT images, the septa and solid component of the mass showed marked enhancement and delay enhancement, while the cystic component showed no enhancement. ② MRI features: Unilateral mass was revealed in 13 cases and bilateral mass was in 1 case. The maximum diameter of the mass ranged from 9.2 to 30.0 cm, mean (14.03±4.72) cm. One mass was irregular, 2 masses revealed incomplete capsule, and 2 masses had septa in the mass. The cystic component showed heterogenous signal intensity on T1WI, and high signal intensity on T2WI. There was no enhancement on post-contrast images. The solid component showed iso-intensity on T1WI, high signal intensity on T2WI, and diffused restricted on diffusion-weighted images. There was markedly enhancement on post-contrast images. ③ Blood supply of the tumor: In 8 cases, the branch of enlarged ipsilateral ovarian artery fed the tumors. In other 16 cases, the masses were surrounded by enlarged ipsilateral ovarian vein. Conclusion The characteristic CT and MRI features of OCCC include: a cystic solid mass with complete capsule; the solid component projects into the cavity, which could be hypervascular and marked enhanced.

Objective To investigate the effect and the mechanism of up-regulating histone acetylizad level with a selective inhibitor of HDACs-Valproate acid sodium (VPA) on breast cancer cell proliferation. Methods MCF-7 cells were cultured with 0.75-4.0 mmol/L valproic acid (VPA) for 24, 48, 72, 96 hours in vitro, the inhibiting rate was tested by MTT assay. Cell cycle was analyzed by flow eytome- try with PI assay, and the protein and mRNA expressions of Cyelin A, Cyclin DI, Cyclin E, P21Waf/cipl of MCF-7 cells after 1.5, 3.0 mmol/ L VPA treated were analyzed by indirect immunofluorescence technique and RT-PCR respectively. Results After cultured with 0.75 -4.0 mmol/L valproic acid (VPA) for 24, 48, 72, 96 hours, the inhibiting rate of experimental groups increased significantly(P<0.01) and a dose and acting time dependent manner was found. As to cell cycle, the percentages of GI, S, M phrase in control groups remained the same. Contrary to control groups, 0. 75 -4.0 mmo]/L VPA induced a significant arrest in G1 phrase ( P<0.01), and a total of 55.4% -82.8% G1 phrase ratio were found. P21Waf/cipl was up-regulated both at the mRNA and protein level while Cyclin D1 was down-regulated ( P<0.001). Conversely, neither mRNA nor protein expression of Cyclin A, Cyclin E showed difference ( P>0.05). Conclusions Up- regulating histone acetylizad level can inhibit breast cancer cell proliferation, induce cell cycle arrest in G1 phrase. VPA, as a I class of histone deaeetylase inhibitor, can be used as an option in the treatment of breast cancer. The mechanism may include up-regulating P21Waf/cipl mRNA and protein expression and down-regulating Cyclin D1 mRNA and protein expression.

Objective: To investigate antisense integrin?V and ?3 gene therapy in Rats mod- el of Pancreatic Carcinoma. Methods: The antisense integrin?V and ?3 expression plasmids wereelectrobloted into pancreatic tumours of rats induced by dimethylbenzanthracine,The tumor inhibit rate was calculated, the tumor microvessel density(MVD)with the immunohistochemical staining and tumor apoptosis with TUNEL were examined. Results: The tumor weight of the control group, ?V group, ?3 group an- d?V?3 group was (1.167?0.79)g(,0.953?0.26)g(,1.013?0.42)g(,0.788?0.56)g respectively. The dif- ference was significant among them(P

Objective:To investigate modulation of a specific HDAC inhibitor,Valproate acid sodium(VPA),on expression of Caspase3,Caspase8,Caspase9 by inhiting HDAC,as well as apoptosis rate of cancer cells treated with VPA and the specific inhibitors of Caspase3,Caspase8,Caspase9.Methods:Heptocellular carcinoma cells-HepG2,gastric carcinoma cells-BGC-823 and breast cancer cells-MCF-7 were cultured with 0.75-4.0 mmol/L of VPA for 48 hrs in vivo,apoptosis was analyzed by flow cytometry with Annexin V/PI assay.The activities and protein expressions of Caspase3,Caspase8,Caspase9 were detected by spectrophotometry and indirect immunofluorescence technique.Results:Contrary to control groups,VPA at concentrations between 0.75 and 4.0 mmol/L induced a significant apoptosis in HepG2,BGC-823,MCF-7 cells(P0.05).The apoptosis rates of cancer cells treated with VPA and specific inhibitors of Caspase3,Caspase9 together was lower than in the groups with VPA treatment singlely(P

PurposeMultiple sclerosis (MS) is characterized by time and spatial multiple, and it is the main reason for disabled young people. This paper aims to investigate the application of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) with dual-compartment Tofts model in relapsing-remitting multiple sclerosis (RRMS) and its correlation with clinical scoring.Materials and MethodsThe clinical data of 25 patients with RRMS were retrospectively studied. The patients underwent the conventional MRI and the DCE-MRI examination. The result was processed by dual-compartment Tofts model and quantitative measurement was carried out in terms of volume transfer constant (Ktrans), rate constant between EES and blood plasma (Kep) and the volume of EES per unit volume of tissue (Ve), cerebral blood flow (CBF) and cerebral blood volume (CBV) of the lesions and normal-appearing white matter (NAWM) regions. The correlation between imaging biomarkers, expanded disability states scale (EDSS) and disease duration were also analyzed.Results ① The differences of MR imaging biomarkers Ktrans and Kep were significant between the regions of nonenhancing (NE) lesions, the NAWM regions near NE lesions and the NAWM regions far from NE lesions (χ2=6.777 and 22.343,P<0.05); however, Ve in the NE lesions had no significant differences compared with that in the NAWM regions near and far from NE lesions (P>0.05).②The CBF and CBV among these three groups had no signiifcant differences (P>0.05).③The CBF of NE lesions was signiifcantly correlated with disease duration (r=0.518,P<0.05);however, the other markers like Ktrans, Kep, Ve, CBF and CBV were neither signiifcantly correlated with EDSS nor with disease duration (r=-0.371-0.052,P>0.05).Conclusion DCE-MRI with Tofts model can quantitatively measure microvascular permeability and perfusion characteristics of lesions and NAWM regions, which thus reflects hemodynamic changes in patients with multiple sclerosis.

Objective To validate the reproducibility of abnormal cerebral metabolic characteristics in PD patients from different medical centers using 18F-FDG PET imaging.Methods A total of 108 subjects who were referred for resting-state brain 18 F-FDG PET imaging were retrospectively reviewed.Thirtythree PD patients (15 males,18 females,age:38-79 years) and 33 age-matched healthy controls (15 males,18 females,age:40-77 years) underwent evaluation at Shanghai Huashan Hospital Affiliated to Fudan University.Seventeen PD patients (10 males,7 females,age:44-74 years) and 17 age-matched healthy controls (6 males,11 females,age:42-67 years) underwent evaluation at the First Affiliated Hospital of Sun Yat-sen University.SPM was used to investigate the cerebral metabolic characteristics of the patients with two-sample t test.Statistically significant voxels were obtained by using familywise error rate (FWE;P＜0.05).Two sets of PD patients with abnormal glucose metabolism of brain regions were obtained and the cerebral metabolic characteristics were compared.Results Regarding the PD patients from Shanghai Huashan Hospital,the features of cerebral glucose metabolism by SPM analysis were demonstrated as follows:increased metabolism was found in the region of pons,cerebellum,thalamus,putamen and pallidum,while decreased metabolism was displayed in the region of parietal lobe and occipital lobe.The increased regions referred to 8 110 voxels and decreased regions referred to 2 810 voxels (P＜0.05).The similar metabolic pattern was found in PD patients from the First Affiliated Hospital of Sun Yat-sen University.The increased metabolism was shown in the regions of pons,cerebellum,thalamus,putamen and pallidum,and referred to 15 573 voxels.The metabolism-decreased regions included parietal lobe,occipital lobe and frontal lobe,and referred to 3 945 voxels (P＜0.05).Conclusions 18F-FDG PET/CT imaging results demonstrate similar metabolic pattern in PD patients from different medical centers,in whom the metabolism-increased regions are found in the pons,cerebellum,thalamus and pallidum and decreased regions were demonstrated in the parietal lobe and occipital lobe.The reproducibility from 18F-FDG PET/CT imaging provides reliable evidence for the multi-center study in the differential diagnosis of PD.

Objective:To explore the effects of Chaihu-Shugan San (CSS) on the behavior and neurogenesis function of depression model mice induced by chronic unpredictable mild stress (CUMS).Methods:Thirty clean grade healthy male C57BL/6 adult mice were randomly divided into control group (Con group), model group (CUMS group) and Chaihu-Shugan San treatment group (CSS group), with 10 mice in each group.The mice in CUMS group and CSS group were given CUMS intervention to establish depression model. At the same time of modeling, the mice in CUMS group and CSS group were given distilled water and CSS(2.7 g/kg) by gavage respectively.While the mice in Con group were only given equal volume distilled water by gavage without CUMS stimulation.After the intervention, the depressive-like behavior of mice was evaluated by increased body weight, sugar water preference test (SPT), forced swimming test (FST) and tail suspension test (TST). The number of newborn neurons was detected by immunofluorescence staining. The mRNA expression levels of brain-derived neurotrophic factor (BDNF), fibroblast growth factor 2 (FGF2) and spindle and kinetochore-associated protein 2(SKA2) in mice hippocampus were detected by qRT-PCR.Statistical analysis was performed by SPSS 22.0 software. One-way ANOVA was used for multi group comparison, and Tukey test was used for pairwise comparison.Results:(1) After modeling, there was significant difference in body weight increment among the three groups ( F=8.859, P <0.05). The body weight increment of CUMS group was lower than those of Con group and CSS group (both P< 0.05). There were significant differences in sugar water preference rate, tail suspension immobility time and swimming immobility time among the three groups ( F=10.544, 12.957, 8.095, all P<0.05). The sugar water preference rate in CUMS group was lower than that in Con group ((87.46±2.78)%, (93.90±3.31)%, P<0.05), and that in CSS group was higher than that in CUMS group ((91.65±2.61)%)( P<0.05). The tail suspension immobility time ((198.00±27.57) s) and swimming immobility time ((322.20±46.98) s) in CUMS group were higher than those in Con group ((138.80±38.50) s, (238.50±50.51) s, both P<0.05). The tail suspension immobility time ((139.00±21.29) s) and swimming immobility time ((265.20±44.90) s) in CSS group were lower than those in CUMS group (both P<0.05). (2) Immunofluorescence showed that there was significant difference in the number of newborn neurons labeled by BrdU and NeuN in the dentate gyrus of hippocampus among the three groups ( F=9.486, P<0.05). The number of double labeled cells (31.66±3.21) in CUMS group was lower than that in Con group(63.66±15.17) and CSS group (58.00±6.00) (both P<0.05). (3) RT-PCR results showed that the mRNA levels of BDNF, FGF2, SKA2 in hippocampal dentate gyrus of the three group were significantly different( F=14.522, 9.337, 8.701, all P<0.05). The levels of BDNF mRNA (0.79±0.06), FGF2 mRNA (0.74±0.18) and SKA2 mRNA (0.52±0.32) in the dentate gyrus of hippocampus in CUMS group were lower than those in Con group (BDNF mRNA (1.03±0.10), FGF2 mRNA (1.04±0.11), SKA2 mRNA (1.05±0.37), all P<0.05). Compared with CUMS group, the mRNA levels of BDNF (1.07±0.80), FGF2 (1.30±0.29) and SKA2 (1.40±0.55) in CSS group were higher (all P<0.05). Conclusion:CSS can alleviate the depressive like behavior of depression model mice, which may be related with increasing the mRNA expression levels of BDNF, FGF2, SKA2 and promoting the proliferation of neural stem cells in hippocampus.

Aim To investigate the effect of theaflavin on oxidized low density lipoprotein(ox-LDL)-induced foam cell formation and oxidative stress in THP-1 macrophages and its mechanism.Methods THP-1 derived macro-phages were pretreated with 50 μmol/L theaflavin and(or)10 μmol/L nuclear factor erythroid 2-related factor 2(NRF2)inhibitor ML385,then 100 mg/L ox-LDL was added to the cells for 24 h to establish the foam cell model.The effect of theaflavin on THP-1 macrophages viability was evaluated by CCK-8 assay and LDH release.The expression of inflamma-tory cytokines interleukin-6(IL-6),interleukin-1 β(IL-1β),tumor necrosis factor-α(TNF-α)were analyzed by real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot.The release of inflammatory cytokines were detected by enzyme linked immunosorbent assay(ELISA).Intracellular lipid accumulation was detected by Oil red O staining,and lipid absorption was observed by DiL-labeled oxidized low density lipoprotein(DiL-ox-LDL)staining.Re-active oxygen species(ROS)level was detected by DCFH-DA probe.The expression of lipid uptake,cholesterol efflux and oxidative stress-related proteins were detected by Western blot and RT-qPCR.Results Treatment with 100 mg/L ox-LDL significantly decreased cell viability and cholesterol efflux-related protein expressions,increased lipid uptake,ac-cumulation and lipid uptake-related protein expressions,and significantly promoted inflammation and ROS level,as well as the expressions of myeloperoxidase(MPO),NADPH oxidase 2(NOX2)in THP-1 macrophages(all P＜0.05).After pretreatment with theaflavin,cell viability was increased,intracellular lipid uptake,accumulation and lipid uptake-related protein expressions were significantly reduced,cholesterol efflux-related protein expressions were significantly increased,the expression and release of IL-6,IL-1β and TNF-α were significantly decreased,ROS level was significantly decreased,and the expression of MPO and NOX2 were decreased(all P＜0.05).Pretreatment with theaflavin effectively alleviated intracellular oxidative stress by altering the expression of NRF2,heme oxygenase-1(HO-1)and Kelch-like ECH-associated protein 1(KEAP1)in NRF2 signaling pathway,and enhanced the translocation of NRF2 into the nucleus.After pretreat-ment with ML385,the expression levels of NRF2,HO-1,KEAP1 and CD36 were significantly decreased.Conclusion Theaflavin can significantly inhibit ox-LDL-induced foam cell formation,inflammation,and oxidative stress through NRF2/HO-1 signaling pathway in THP-1 macrophages.