Objective To explore the effects of murinecytomegalovirus(MCMV)infected mouse embryonic fibroblasts(MEF)on the proliferation and activation of regulatory T cell in vitro. Methods A co-culture system of T cell and MCMV infected MEF(T-MEF MCMV)was established.The viral load of supernatant was determined by plaque assay.The proliferation of T cells was observed with cell counting.The proportion of CD4+ CD25+ cells was measured by flow cytometry.The levels of Foxp3 protein were measured by Western blot.Reverse transcription polymerase chain reaction(RT-PCR)assay was utilized tO determine whether MCMV infection of MEF influenced the level of transforming growth factor(TGF)-β mRNA.The level of TGF-β protein in supernatant was measured by double-antibody sandwich enzyme linked immunosorbent assay(ELISA).The difference between T-MEF MCMV group and control group was assessed by one-way ANOVA.Results When T cells were co-cultured with MCMV infected MEF for 1 day and 3 days,the viral load in supernatant decreased.But when co-culture lasted for 6 days,the antiviral effect obviously diminished,as the viral load[(5.58±0.67)×105 PFU/mL]of the experimental group showed no statistic difference with MEF MCMV control group[(6.05±0.34)×105PFU/mL].When co-cultured with MCMV infected MEF for 3 days,T cell increased from pre-culture level of[(2.02±0.05)×106/mL]to(2.25± 0.13)×106/mL(P＜0.05).But when co-culture lasted for 6 days,the number of T eelI returned to (2.08±0.14)×106/mL,which had no statistic difference with that of co-culture for 3 days system. Both the expressions of Foxp3 protein and the proportion of CD4+ CD25+ Foxp3+ Treg cells in T-MEF MCMV group were up-regulated as the infection time extended,which were two times and three times of the control group level,respectively.The mRNA level(A value)of TGF-β in MCMV infected MEF increased from baseline of 1.09±0.13 to 3.15±0.54 on day 3 after infection.The expression of TGF-β in supernatant 3 days after infection was(3.85±0.32) μg/L,which was significantly higher than that before infection[(1.74±0.14)μg/L,P＜0.05].Conclusions Activated T cells have antiviral effect.However,the function of T cells is rapidly inhibited after activation,which may be due to the expression of Foxp3 mRNA induced by MCMV infected MEF and increased CD4+ CD25+ Treg proportion of the co-cultured T cells.TGF-β level is significantly increased after CMV infection,which may be an important mechanism of Treg proliferation.MCMV may manipulate Treg to evade specific immune elimination and,as a result,to cause CMV replication.

OBJECTIVE: To study chemical constituents from the twigs and leaves of Illicius majus Hoot et Thoms. METHODS: The compounds were isolated by column chromatography on silica gel, Sephadex LH-20, and flash chromatography coupled with preparative HPLC. Their structures were elucidated by spectroscopic analysis including IR, NMR, and MS. RESULTS: Six compounds, ie. majusanicol(1), angustanoic acid G(2), 4-hydroxyphenethyl alcohol(3), 4-hydroxybenzoic acid(4), 3-hydroxybenzoic acid(5), and 3, 5-dimethoxy-4-hydroxybenzoic acid(6), were isolated from the twigs and leaves of I. majus. CONCLUSION: Compound 1 is a new abietane-type diterpenoid, and compound 3-6 are isolated from this plant for the first time.

Objective: To study the chemical constituents from the twigs and leaves of Illicius majus. Methods: The compounds were isolated by column chromatography over silica gel, Sephadex LH-20, and flash chromatography coupled with preparative HPLC. The structures were elucidated by spectroscopic analysis including ESI-MS, 1D-NMR, and 2D-NMR. The cytotoxic activities were assessed by MTT assay. Results: Compounds 1—14 were isolated from the twigs and leaves of I. majus, and identified as 3′-methoxy-kaempferol-3-O-α-L-rhamnopyranoside (1), quercetin-3-O-arabinoside (2), kaempferol-3-O-arabinoside (3), 8′-oxo-6-hydroxy-dihydrophaseic acid (4), 4-O-methylcedrusin (5), (−)-massoniresinol (6), (7S,8R)-3,3′,5-trimethoxy-4′,7-epoxy- 8,5′-neolignan-4,9,9′-triol (7), vladinol F (8), 2,3-dihydro-2-(4′-α-L-rhamnopyranosyl-3′-methoxyphenyl)-3-hydroxymethyl-7- methoxy-5-benzofuranpropanol (9), quercetin-3-O-α-L-rhamnopyranoside (10), kaempferol-3-O-α-L-rhamnopyranoside (11), vitexin (12), 3,5,7-trihydroxychromone-3-O-glucopyranoside (13), and benzyl alcohol O-β-D-glucopyranoside (14). Compounds 1—14 exhibited the cytotoxic activity against HCT-8, Bel-7402, BGC-823, and A549 with IC50 values of over 10 μmol/L, respectively. Conclusion: Compounds 1—14 are all obtained from I. majus for the first time, and compounds 2, 3, 5, 7, 8, 9, and 14 are obtained from the genus of Illicium for the first time. All compounds show no significant cytotoxic activities against HCT-8, Bel-7402, BGC-823, and A549 cancer cell.

Objective: To study the chemical constituents from the twigs and leaves of Illicius majus. Methods: The compound was isolated by column chromatography over silica gel, Sephadex HL-20, and flash chromatography coupled with preparative HPLC. The structure was elucidated by spectroscopic analysis including HR-ESI-MS, 1D-NMR, and 2D-NMR. The cytotoxic activities were assessed by MTT. Results: A new seco-prezizaane-type sesquiterpenoid named majusatone (1) was isolated from the twigs and leaves of I. majus. Compound 1 exhibited the cytotoxic activity against HCT-8, Bel-7402, BGC-823, and A549 with IC50 values of 42.3, 48.5, 51.1, and 39.9 μmol/L, respectively. Conclusion: Compound 1 is a new special seco-prezizaane-type sesquiterpenoid, and shows no significant cytotoxic activities against HCT-8, Bel-7402, BGC-823, and A549.

OBJECTIVETo study the changes of biomarkers in cerebrospinal fluid (CSF) in cerebral amyloid angiopathy (CAA) dementia and Alzheimer(')s disease.

METHODSLevels of amyloid protein β (Aβ42, Aβ40) and phosphorylated Tau-protein (P-tau) in CSF and ratio of Aβ42/Aβ40 were tested in 5 cases with CAA dementia and 20 cases with Alzheimer's disease collected at Peking Union Medical College Hospital from December 2001 to March 2011.

RESULTSThe levels of Aβ42, Aβ40, and P-tau in CSF and ratio of Aβ42/Aβ40 were (660.4 ± 265.2) ng/L, (7111.0 ± 1033.4) ng/L, (71.8 ± 51.5) ng/L, and 0.077 ± 0.033, respectively in CAA dementia and (663.6 ± 365.6) ng/L, (5115.0 ± 2931.1) ng/L, (47.7 ± 38.8) ng/L, and 0.192 ± 0.140, respectively in Alzheimer's disease patients. There were no statistically significant differences between CAA dementia and Alzheimer's disease in terms of these CSF biomarkers (all P>0.05).

CONCLUSIONMeasurements of CSF biomarkers may not be helpful in differential diagnosis of CAA and Alzheimer's disease.

Aged ; Aged, 80 and over ; Amyloid beta-Peptides ; cerebrospinal fluid ; Apolipoproteins E ; genetics ; Biomarkers ; cerebrospinal fluid ; Cerebral Amyloid Angiopathy ; cerebrospinal fluid ; Dementia ; cerebrospinal fluid ; Humans ; Male ; tau Proteins ; cerebrospinal fluid

This research explored the synergistic effects and the potential mechanisms of RCE-4 and various nonsteroidal anti-inflammatory drugs (NSAIDs) on the proliferation of cervical cancer Ca Ski cells. The MTT assay and CalcuSyn V2.0 software were used to detect cell proliferation and calculate the combination index (CI); the expression levels of various proteins were analyzed using Western blot assay; mitochondrial membrane potential (MMP) was assessed using JC-1 staining; acridine orange/ethidium bromide (AO/EB) double-fluorescence staining was used to detect the apoptosis of Ca Ski cells; a co-immunoprecipitation (Co-IP) assay was used to analyze the relative content of Bcl-2-Beclin 1 complex in Ca Ski cells. The results demonstrate that the combination of RCE-4 and NSAIDs increases the inhibition of Ca Ski cells compared to the single-RCE-4 group, and celecoxib provided the best synergistic effect among the four NSAIDs tested, with a CI of 0.32. The combination of RCE-4 and celecoxib significantly down-regulated the expression of cyclooxygenase-2 (COX-2) and nuclear transcription factor-κB (NF-κB), and promoted the expression of non-steroidal anti-inflammatory drugs activity gene-1 (NAG-1). In addition, autophagy induced by RCE-4 was markedly inhibited in combination with celecoxib, which was associated with down-regulation of the expression of microtubule-associated protein 3 (LC3)-II, Beclin 1, p62 and autophagy-related gene (ATG) 3/4B/5/7/14. RCE-4-induced apoptosis was significantly enhanced by altering the depolarization of mitochondrial membrane potential and the expression of B cell lymphoma-2 (Bcl-2), B cell lymphoma-xl (Bcl-xl), Bcl-2 associated X protein (Bax), Bcl-2/Bcl-xl-associated death promoter (Bad) and cleaved cysteinyl aspartate specific proteinase (cleaved-caspase) 3/7/9. Furthermore, the formation of the Bcl-2-Beclin 1 complex was significantly inhibited in Ca Ski cells treated with RCE-4 in combination with celecoxib. Taken together, this research shows that the combination of RCE-4 and celecoxib has a significant synergistic effect on the proliferation of Ca Ski cells by promoting apoptosis, inhibiting autophagy and disturbing the formation of the Bcl-2-Beclin 1 complex, which may be a novel strategy to increase the sensitivity of anti-cervical cancer drugs.

Adrenalectomy ; Adult ; Angiomyolipoma ; pathology ; surgery ; Follow-Up Studies ; Humans ; Kidney ; pathology ; Kidney Neoplasms ; pathology ; surgery ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Nephrectomy ; Ureter ; surgery

Objective To explore the role of CD4+CD25+regulatory T cells(Treg)in the co-culture svstem consisting of T cells and mouse embryo fibroblasts(MEF)infected with murine cytomegalovirus (MCMV)in vitro.Methods A co-culture system of T cells and MCMV infected MEF(MEFMCMV)was established.The viral load in the supernatants was determined by plaque assay.TH1/TH2 differentiation-specific transcription factors T-bet/GATA-3 were assayed by Western blot.The levels of cytokines IL-4,IL-10and IFN-γ in the co-culture supenatants were measured by double-antibody sandwich ELISA.Results After 3 d incubation,the viral load in the supernatants of TdepTreg-MEFMCMC group,in which the T cells depleted Treg(TdepTreg)were co-cultured together with MEFMVMV,was significantly lower than that in MEFMCMV group.And in the co-cultivation of MEFMCMV and T cells without Treg the expressions of T-bet/GATA-3,IL-4,IL-10 and IFN-γ incteased significantly.Compared with the virus content in the TdepTreg-MEFMCMV group,the MCMV load in the"add-on Treg group"increased and the levels of T-bet/GATA-3 and IFN-γ were lower,and the expression of IL-4 didn't show any significant difference. Compared with the level of IL-10 in the TdepTreg-MEFMCMV group,the IL-10 level in the"add-on TREG group"didn't show any significant differece when the Treg ratio among total T cells was 1%～2%,and increased significantly when the Treg ratio was more than 5%.These eflfects were all correlated with Treg ratios in a dose-dependent manner.Conclusion MCMV infected MEF can induce the proliferation and activation of effector T cells,but Treg can inhibit the T cell-mediated protective effect on MCMV infection.

Objective To study the expression of intermediate-conductance-Ca~(2+)-activated K~+ (IKCa1)channels in endometrial cancer and its role in regulating proliferation of endometrial cancer cells. Methods Western blot and RT-PCR were used to examine the expression of IKCa1 channels in 13 normal endometrial specimens and 25 endometrial cancer specimens;and RNA interference(RNAi),[ ~3H ] thymidine incorporation,and inhibitor of IKCa1 channel were used to explore the role of IKCa1 channels in regulation of proliferation of endometrial cancer cells HEC-1A.Results The expression rate and level of IKCa1 mRNA in endometrial carcinoma(84%,0.89?0.52)were higher than in normal endometria(8%, 0.14?0.12;P

Aim To explore the effect of receptor component protein(RCP)in the signal transduction of vascular smooth muscle cell(VSMC)proliferation induced by static pressure.Methods The mouse-derived vascular smooth muscle cell line(A10VSMC)was employed in the experiment.Cells were exposed to static pressure,and MTT assay was used to detect the cell viability.Western blot was used to determine the expressions of PCNA,RCP and p-Akt,RCP mRNA was tested by RT-PCR,and co-immunoprecipitation was used to test the interaction between RCP and G proteins.Results The cell viability,expressions of PCNA and RCP increased with the elevation of static pressure and reached their peaks at 120 mmHg,and after 6 hours they got a plateau.The static pressure significantly increased the level of p-Akt,meanwhile,the binding of RCP and Gαs significantly decreased.However,the binding of RCP and Gβ increased in response to static pressure after stimuli of static pressure,but Gγ was obscure.Conclusion Static pressure can induce VSMC proliferation and expression of RCP,which may involve G protein signal transduction model.