Objective To study the immunoadjuvant effects of chitosan oligosaccharide(COS),including the immune activation and the triggering of lysosomal escape,and to explore whether COS can be used as an adjuvant for attenuated live bacteria vector vaccines.Methods 1)Mouse macrophages RAW264.7 cells were cultured with COS at 0 mg/mL(the control group)and 0.1-4 mg/mL for 24 h and the effect on cell viability was measured by CCK8 assay.Mouse macrophages RAW264.7 were treated with COS at 0(the control group),1,2,and 4 mg/mL for 24 h.Then,the mRNA expression levels of the cytokines,including IFN-γ,IL-10,TGF-β,and TLR4,were determined by RT-qPCR assay.2)RAW264.7 cells were treated with 1 mL of PBS containing different components,including calcein at 50 μg/mL,COS at 2 mg/mL,and bafilomycin A1,an inhibitor,at 1 μmol/mL,for culturing.The cells were divided into the Calcein group,Calcein+COS group,and Calcein+COS+Bafilomycin A1 group accordingly.Laser scanning confocal microscopy was used to observe the phagocytosis and the intracellular fluorescence distribution of calcein,a fluorescent dye,in RAW264.7 cells in the presence or absence of COS intervention to determine whether COS was able to trigger lysosomal escape.3)LM?E6E7 and LI?E6E7,the attenuated Listeria vector candidate therapeutic vaccines for cervical cancer,were encapsulated with COS at the mass concentrations of 0.5 mg/mL,1 mg/mL,2 mg/mL,4 mg/mL,and 8 mg/mL.Then,the changes in zeta potential were measured to select the concentration of COS that successfully encapsulated the bacteria.Phagocytosis of the vaccine strains by RAW264.7 cells was measured before and after LM?E6E7 and LI?E6E7 were coated with COS at 2 mg/mL.Results 1)CCK8 assays showed that,compared with the findings for the control group,the intervention of RAW264.7 cells with COS at different concentrations for 24 h was not toxic to the cells and promoted cell proliferation,with the difference being statistically significant(P＜0.05).According to the RT-qPCR results,compared with those of the control group,the COS intervention up-regulated the mRNA levels of TLR4 and IFN-γ in RAW264.7 cells,while it inhibited the mRNA expression levels of TGF-β and IL-10,with the most prominent effect being observed in the 4 mg/mL COS group(P＜0.05).2)Laser scanning confocal microscopy revealed that the amount of fluorescent dye released from lysosomes into the cells was greater in the Calcein+COS group than that in the Calcein group.In other words,a greater amount of fluorescent dye was released from lysosomes into the cells under COS intervention.Furthermore,this process could be blocked by bafilomycin A1.3)The zeta potential results showed that COS could successfully encapsulate the surface of bacteria when its mass concentration reached 2 mg/mL.Before and after the vaccine strain was encapsulated by COS,the phagocytosis of LM?E6E7 by RAW264.7 cells was 5.70%and 22.00%,respectively,showing statistically significant differences(P＜0.05);the phagocytosis of LI?E6E7 by RAW264.7 cells was 1.55%and 6.12%,respectively,showing statistically significant differences(P＜0.05).Conclusion COS has the effect of activating the immune response of macrophages and triggering lysosomal escape.The candidates strains of coated live attenuated bacterial vector vaccines can promote the phagocytosis of bacteria by macrophages.Further research is warranted to develop COS into an adjuvant for bacterial vector vaccine.

Objective To investigate the prevalance and risk factors of post-stroke depression(PSD)in patients with the first ever stroke during first year after stroke.Methods A total of 60 eligible stroke patients were followed up for 12 months after the first ever stroke and Hamilton Rating Scale for Depression,Activity of Daily Living Scale,Neurological Functional Deficit Scores,Social Support Scale and Eysenck Personality Questionnaire(EPQ)were used to investigate the incidence and risk factors of PSD in 2 weeks,3,4 and 12 months,respectively.Results The prevalence of PSD after the first ever streke was 18.18%(12/66),29.23%(19/65),38.09%(24/63),31.67%(19/60)in 2 weeks,3,4 and 12 months after the attack of stroke,respectively.Logistic regression analysis indicated that the major risk factors of PSD were female and introvert(b=-1.55,P=0.001;b=1.59,P=0.047)in 2 weeks,poor social support,low grade of neurological functional deficit score and female(b=-0.19,P=0.005;b=0.26,P=0.01;b=2.84,P=0.03)in 3 months.poor social support and low grade of neurological functional deficit score in 6 months and 12 months after stroke re-spectively(6 months:b=-0.24,P<0.001;b=0.35,P=0.004.12 months:b=-0.17,P=0.001;b=0.33,P=0.002).Conclusions The prevalence of PSD is considerable and the risk factors of PSD are different in different stage.

BACKGROUND: Artificial nerve grafts of human hair keratin are a kind of biological products. It has low antigenicity,absorbability and stimulation to nerve fiber growth following specific biochemistry. It is hoped to have better effect than otherartificial nerve grafts.　 OBJECTIVE: To investigate the anatomy and histocompatibility of artificial nerve grafts of the human-hair keratin, and toobserve its effects on the repair of peripheral nerves. 　 DESIGN, TIME AND SETTING: Randomized, controlled, animal experiment. The study was performed at the Animal CentralLaboratory of Affiliated Hospital of Qingdao University Medical College between November 2006 and June 2008. MATERIALS: Artificial nerve grafts of human hair keratin is a compound of human hair processed by specific controlledbiochemistry based on ground substance, embedded with a layer of biological membrane. It has low antigenicity, absorbabilityand stimulation to nerve fiber growth following specific biochemistry. METHODS: Eighteen Wistar rats were randomly divided into three groups. The sciatic nerve, 10 mm, was removed andtransplanted with human-hair keratin graft, skeletal muscle and untreated hair, respectively.MAIN OUTCOME MEASURES: The characteristics of histomorphology and anatomy were observed at 8, 12, 24 weeks afterthe surgery. RESULTS: White tissues appeared between the broken ends of the sciatic nerve at 8 post-operative week in the graft group,and appeared in the graft space in human-hair keratin at the 12th week. At the 24th week, a large amount of infantile myelinatednerve fibers were observed under optical microscope regenerating around the human hair, which was partially degraded andabsorbed. Schwann cells were observed under an electron microscope and myelinization. 　 CONCLUSION: The artificial nerve grafts of the human-hair keratin are well compatible with the body tissues, and couldinduce nerve regeneration.

Objective To explore the relationship between the levels of Cu, Fe, Mn, Zn in serum and Parkinson disease (PD). Methods A total of 40 patients with PD (PD group) and 40 control subjects (control group)were enrolled in this study. Serum levels of Mn were measured by graphite atomic absorption, and Cu ,Fe, Zn were measured by inductively coupled plasma(ICP)mass spectrometry. Resudts There were significantly increase in the levels of Mn and Fe in PD group than those in control group [(0.269±0.326) μ mol/L vs (0.125±0.054) μmol/L, P< 0.05, (1.512±0.949) μmol/L vs (0.676±0.111) μmol/L, P< 0.01)]. There were no significant difference in the levels of Cu and Zn between two groups (P> 0.05). Condusion Micreelements may play important roles in pathogenesis and development of PD, especially Fe and Mn.

Objective To observe the effects of low frequency electrostimulation(LFES)on sleep disorder of patients after acute cerebral infarction(ACI)as evaluated by using polysomnography,and on the recovery of neurological deficits. Methods Seventy cases of acute cerebral infarction were randomly divided into two groups,a treatment group and a control group.Both groups were treated with routine drugs, and the treatment group was also treated with LFES in addition.The changes of neurological deficits(ND) scores and such parameters of polysomnography as sleep latency(SL),total sleep time (TST),sleep efficiency(SE%),sleep maintenance(SMT),rapid eye movement sleep(REM),REM latency(RL),REM time(RT),REM activity(RA),REM density(RD).stage 1 sleep(S1),stage 2 sleep(S2)and deep sleep (S3+4) were observed.Results It was shown that,after treatment,both groups got significant improvement in terms of the ND scores and all the polysomnography parameters except RA,S1 and S2,(P<0.01),and the treatment group improved to a significantly greater extend when compared with the control group (P<0.01).Conclusion It is concluded that LFES could promote recovery of neural function and sleep disorder of ACI patients.