Natural killer ( NK) cells are important innate effector cells and play a vital role in maintaining homeostasis through potent cytotoxic activity and cytokine production. Recent findings show that NK cells can also shape adaptive immune responses by in-fluencing a variety of immune cells. In addition to direct interactions with other immune cells,NK cells can indirectly stimulate or inhibit adaptive immune response via influencing infected cells and pathogen load. Abundant studies have highlighted the positive regulatory functions of NK cells, while their negative regulatory functions have increasingly attracted attention in recent years. Here, we review recent findings on negative regulation of adaptive immune response by NK cells, discussing the involved effector cells and function mechanism,and demonstrate how this negative regulation influences the overall outcome of adaptive immunity in infection and tumor disease.

Objective To study antidepressant effect and mechanism of different polar fraction of Acorus tatarinowii Schott.on depression mouse model and explore the mechanism. Methods Different polar fractions were prepared by systematic solvent method. The adult dose recommended by Chinese Pharmacopeia ( 10 g?d-1 ) was converted to the dose of mouse. Intragastric administration was performed.A total of 80 male mice was randomly divided into normal control group,model control group and different polar fraction groups of Acorus tatarinowii Schott.. Depression mouse model was established by chronic unpredictable mild stimulation ( CUMS) with solitary feeding,and external performance of mice of different groups was observed. After 21-day feeding,blood was harvested from eyes of the mice.Concentrations of triiodothyronine (T3),tetraiodothyronine (T4), thyroid-stimulating hormone ( TSH) ,adrenocorticotrophic hormone ( ACTH) in mouse plasma and 5-hydroxy tryptamine ( 5-HT) , ACTH in brain were measured by ELISA method. Results Concentrations of T3 , T4 and ACTH in model control group [(1.203±0.042),(44.80±2.21),(11.27±0.50) ng?mL-1] were significantly increased as compared with the normal control group [(0.794±0.028),(24.87±1.25),(7.04±0.24) ng?mL-1] (P<0.05).Concentration of 5-HT in brain (146.87±10.96) was significantly decreased as compared with that of normal group ( 237. 11 ± 21. 87 ) ng?L-1 , and concentration of ACTH (58.94±4.46) ng?L-1 was significantly increased as compared with that of normal group [(38.89±2.26) ng?L-1,P<0.05)]. Concentrations of T3[(0.824±0.067),(0.812±0.051),(0.943±0.049) ng?mL-1],T4[(25.97±1.96),(27.53±1.88), (31.26±1.97) ng?mL-1] and ACTH [(7.21±0.40),(7.58±0.39),(8.69±0.42) ng?mL-1] in extraction group,cyclohexane group and chloroform group were significantly decreased as compared with model control group. At the recommendation dose of Chinese Pharmacopeia,concentrations of 5-HT in brain [(219.59±10.48),(202.19±11.95),(186.96±10.29) ng?L-1] were significantly increased,and concentrations of ACTH [(41.65±2.65),(44.87±2.95),(47.75±3.06) ng?L-1] were decreased ( P<0.05) . Conclusion Mouse model of CUMS shows typical depression symptom,and the symptom has relationship with the concentrations of endocrine hormones of T3 , T4 , ACTH, 5-HT, etc. Extraction and low polar fractions ( cyclohexane, and chloroform) of Acorus tatarinowii Schott. can partly correct incretion disorder of depression model, which may be parts of the reasons why Acorus tatarinowii Schott.exerts anti-depressant effect.

Based on the standards for mental developmental re-tardation set up by WHO in 1985,by applying IQ andadaptation to social behavior as indexes for observa-tion with testing methods for intelligence commonlyaccepted,we have observed the effects in 128 suchcases treated by combined use of acupuncture,adhe-sive—pressing therapy at auricular points and drugplastering at points.Results showed that our therapyis conducive to the improvement of intelligence in suchcases.

Objective To investigate the influence of murine cytomegalovirus(MCMV) infection on differentiation and differentiation gene expression of neural stem cells (NSCs) in vitro for studying the mechanisms of brain abnormalities calmed by congenital cytomegalovirns infection. Methods NSCs were separated from fetal BALB/c mouse and cultured and identified in vitro. The differentiation potency of NSCs was observed by immunnfluorescence. The NSCs infected by MCMV at dosage of multiplicity of infection (MOI) equaled to 5, I and 0. 1, respectively, were cultured in differentiation medium. The morphological changes of the cells were observed by inverted microscope. The ratios of NSCs and its differentiated cells were detected by flow cytometry. The expression changes of nestin, GFAP and NSE, markers of NSCs and its differentiated cells, were studied by immunofluorescence ( MOI = 1 ). The expression of early antigen (EA) of MCMV was detected to observe the infection process. Real-time RT-PCR method was employed to measure the expression levels of the key differentiation genes Wnt-3 and Wnt-7a in Wnt signal pathway of NSCs at early phage of differentiation culture. Results NSCs isolated from embryonic mouse brains could proliferate to form neurnspheres and strongly express Nestin and differentiate into NF-200 positive neurons or GFAP positive astrocytes. The NSCs of the infected groups couldn't adhere to the wall and appear differentia-tion growth, but showed swollen gradually after differentiation culture. The nostin expression of the infected groups downregulated slowly and was higher than that of the control groups ( P < 0.05 ). The GFAP and NSE expression of the infected groups were lower than that of the control groups (P <0.05). The EA of MCMV could be always detected in the cells of the infected groups. The ratios of nestin positive cells of the infected groups were higher than that of the control groups, but the ratios of GFAP and NSE positive cells of the for-mer were lower than that of the latter from 3rd to 9th day after differentiation culture ( P < 0.05 ). The levels of Wnt-3 mRNA and Wnt-7a mRNA of the infected groups were markedly lower than that of the control groups from 1st to 2nd clay and from 12th hour to 2nd day after differentiation culture respectively ( P < 0.05 ) . These changes of the infected groups became more obvious as MCMV MOI increased . Conclusion MCMV could inhibit significantly NSCs differentiate to neurons and astrocytes and lead to the decrease of dif-ferentiated cells. MCMV could inhibit or interfere with the gene expression of Wnt-3 and Wnt-7a in Wnt sig-nal pathway of NSCs. The effect that MCMV inhibited the differentiation and the differentiation gene expres-sion of NSCs showed dose-dependent with MCMV MOI. The inhibitory effect of MCMV on the differentiation of NSCs might be induced by interfering the differentiation gene expression of NSCs, which is possibly the one of primary causes of brain development disorders caused by congenital CMV infection.

To study the effect of endotoxin on liver apoptosis, L02 liver cells were cultured and passaged in vitro, and then stimulated by endotoxin at 10 mg/mL for 4, 8, 16 and 24 h respectively. Liver apoptosis was flow cytometrically and fluorescently detected. Immunohistochemistry was used to detect the delivery of smac and caspase9. The delivery of liver cell smac and the activity of caspase3 were measured by caspase3 assay kit. The hepatic failure models of rats were established by using D-galactosamine. The blood serum and liver tissues were collected for the detection of the liver function, the level of endotoxin and the activity of caspase3 by using chromogenic substrate limulus amebocyte lysate method (LAL) and caspase3 active assay kit. The expression of smac and caspase9 in liver cells was detected by Western blotting. With in vitro study, the L02 cells stimulated by LPS condensed into conglobation and formed apoptotic bodies. After those cells were stained by hoechst, the apoptotic cells displayed blue color under the fluorescent microscope. The apoptosis rate was increased over time and the apoptosis was mainly of advanced stage. Meanwhile, the rate of smac delivery and activity of caspase9 and caspase3 were increased on L02 cell membrane. In vivo, hepatic failure and obvious endotoxemia were induced by injection of more than 200 mg/kg D-GalN. Hepatic mitochondria smac was reduced with dosage of D-GalN and, on the contrary, the activity of caspase3 was increased. D-GalN at 200 mg/kg increased Caspase9 while D-GalN at 300 mg/kg decreased caspase9. Mitochondria signal channel plays an important role in the endotoxin-induced apoptosis of hepatic cells by promoting the release of smac from mitochondria to cytoplasm and activating caspase9 and caspase3 in its low-level channel.
Apoptosis/*drug effects ; Carrier Proteins/*metabolism ; Caspase 3/metabolism ; Caspase 9/metabolism ; Cell Line ; Cells, Cultured ; Endotoxins/*pharmacology ; Liver/cytology ; Liver/*metabolism ; Liver/pathology ; Liver Failure/chemically induced ; Liver Failure/pathology ; Mitochondrial Proteins/*metabolism ; Rats, Wistar

Objective To investigate the biological function of M122 in pathogenesis of MCMV in developmental brain disorders and brain damage, screening for mouse brain cDNA library interacting with M122 was performed by a yeast two-hybrid system. Methods The reconstructed bait plasmid pGBKT7-M122 was transformed into yeast cells AH109 and screened on the nutrient deficiency medium SD/-Trp. After express of the bait protein in AH109 yeast strains was detected by Western blot analysis, yeast-two hybrid screening was performed by mating AH109 with Y187 containing mouse brain cDNA library plasmid. The diploid yeast cells were plated on the nutrient deficiency medium SD/-Trp/-Leu/-His/-Ade. The second screening was performed with SD/-Trp/-Leu/-His/-Ade containing X-α-gal. The plasmids in positive colonies were extracted and transformed into E. coli JM109 cells. After plasmid DNA in JM109 cells were extracted form positive colonies and sequenced, the results were analyzed by bioinformatic methods. The interactions between M122 protein and the protein obtained from positive colonies were further confirmed by repeating yeast-two hybrid. Then, autoactivations of the proteins obtained from positive colonies were detected.Results The reconstructed bait plasmid was transformed into yeast cells AH109 successfully. The bait protein expressed in the yeast cells AH109 stably. 24 proteins interacting with MCMV M122 were screened, including syntaxin 8 ( Stx8 ), phosphoglucomutase 2 ( Pgm2 ), potassium voltage-gated channel, shaker-related subfamily, beta member 1 ( Kcnab1 ), collagen, type ⅪⅩ, alpha 1 ( Col19a1 ), archain 1 ( Arcn1 ), cytidylate kinase( Cmpk), DnaJ(Hsp40) homolog, subfamily A, member 1 (Dnaja1), ATPase, Na+/K + transporting, beta 3 polypeptide( Atp1b3 ), SH3-domain GRB2-like ( endophilin ) interacting protein 1 ( Sgip1 ),ankyrin repeat domain 17 (Ankrd17), Smg-7 homolog, nonsense mediated mRNA decay factor(Smg7),sperm associated antigen 9 ( Spag9 ), FK506 binding protein 1a ( Fkbp1a), MYST histone acetyltransferase monocytic leukemia 4 ( Myst4), hyaluronan and proteoglycan link protein 1 ( Hapln1), autophagy-related 3 (Atg3), splicing factor, arginine/serine-rich 5 ( Sfrs5 ), zinc finger, C3HC-type containing 1 ( Zc3hc1 ),thioredoxin-related transmembrane protein 1 ( Txndc1 ), adaptor protein complex AP-1, gamma 1 subunit (Ap1g1), Cullin 1 ( Cul1 ), and so on. Three of them were formerly unknown proteins. M122 protein could interact with the proteins obtained from positive colonies in the yeast cells AH109. Ap1g1 and Cul1 were proved to have autoactivation. Conclusion A class of proteins in brain interacting with M122 has been obtained. It is presumed that these proteins are correlated with neuropathogenesis of the brain disorders caused by CMV, but the candidates still need further confirmation for the interaction.

Objective To explore the relationship between methylenetetra hydrofolate reduetase (MTHFR) C677T genotypo and unstable angina pectoris(UA) in Chinese population. Methods The study consisted of 90 UA cases (UA group), and an age- and sex- matched healthy control cases (control group, n = 90). PC R-RFLP was used to analyze polymorphism of the MTHFR C677T genotypo. The relationship between MTHFR C677T genotype and UA was observed. Results MTHFR 677C→T mutation was found in 30 of 90 patients with unstable angina pectoris (33.33%) and in 15 of 90 control subjects (16.67%). This difference was statistically significant (P<0.05). Conclusion MTHFR 677C→T mutation is closely related to the unstable angina poctoris.

Objective:To investigate the characteristics of health-related risky behaviors in adolescents with bipolar disorder.Methods:Fifty adolescents aged 12 -18 years,fulfilled the International Statistical Classification of Diseases and Related Health Problems,Tenth Revision(ICD-10)criteria for bipolar disorder(29 in depressive state, 19 in manic state,and 2 in mixed state)and 100 healthy-matched controls were recruited.Their health-related risky behaviors were assessed using the Questionnaire for Adolescents Health-related Risky Behavior Inventory (AHRBI) and the Questionnaire for Adolescents Health-related Risky Behavior Inventory for Parent (AHRBI-P).Results:The bipolar group had higher AHRBI scores in the total scale and six subscales than controls,including Aggression and Violence (AV),Health-Compromising Behavior (HCB),Rule Breaking (RB),Unprotected Sex (US),Self-injury and Suicide (SS),and Smoking and Drinking(SD)[Total scores,55.5(38,119)vs.46(38,65);P <0.05].Besides, the scores of 26 items of bipolar group were all higher than controls(Ps <0.05).The scores in the total scale and six subscales had no difference between AHRBI and AHRBI-P.According to the partial correlation analysis (de-pressive state =0;manic state =1),the AHRBI and AHRBI-P scores of Self-injury and Suicide subscale were nega-tively associated with the episode state (r =-0.32,-0.33;Ps <0.05).The AHRBI scores of'destroy properties'which belongs to the Aggression and Violence subscale were positively associated with the episode state (r =0.32, P <0.05).Conclusion:The adolescents with bipolar disorder have more health-related risky behaviors than the healthy adolescents.The depressive patients have higher risk of suicide.While,the risky behavior of destroying properties tend to occur among the manic patients.

Objective To investigate the effect of stromal cell-derived factor-1α( SDF-1α) and interleukin ( IL-1β) on inducing vascular endothelial cells to express lymphatic phenotype .Methods The CRL-1730 cell line was cultured and treated with SDF-1αor IL-1β.The expression of endothelial cell markers and lymphatic endothelial cell markers were investigated with Real-time PCR, Western blotting and immunocytochemistry .Results In CRL-1730 cell line, endothelial cell markers such as voln willebrand factor ( vWF ) , VE-cadherin , vascular endothelial growth factor receptor(VEGFR)2, were dose dependently down-regulated after SDF-1αstimulation, while lymphatic phenotypes such as Prox-1, podoplanin and lymphatic vessel endothelial hyaluronan receptor-1(LYVE-1), were dose-dependently up-regulated after SDF-1αstimulation.The changes of vWF, VEGFR2 and podoplanin, Prox-1, LYVE-1 expression after IL-1βstimulation was similar to that after SDF-1αwhile expression of VE-cadherin changed slightly .Conclusion SDF-1αand IL-1βare able to induce vascular endothelial cell expressing lymphatic phenotype .

Objective To study the mechanism of RING finger protein 34 ( RNF34 ) involved in innate immunity . Methods Recombinant PCR was used and transient expression of the plasmid was achieved in HEK 293T cells.The cells were stimulated with Sendai virus ( SeV) or N-RIG-Ⅰfor the indicated time while luciferase activity was observed using the dual-luciferase reporter assay kit .Results We constructed the plasmid pcDNA 3-Flag-RNF34 and its three mutations .The study found that when stimulated by SeV , RNF34 could inhibit the activity of NF-κB and IFN-βmore significantly than RNF34-ΔFYVE, RNF34-ΔCID and RNF34-ΔRING.We also found that RNF 34 and its three mutants had similar inhibitory effect when the activation of NF-κB and IFN-βwas stimulated by the N-RIG-Ⅰ.Conclusion RNF34 negatively regulates innate immunity by acting on the RIG-Ⅰ-MAVS signaling pathway .