Objective:To separate and purify the polysaccharides from Polygonatum filipes,characterize their primary structure and investigate the immunomodulatory effects on RAW264.7 macrophages.Methods:Crude polysaccharides from Polygonatum filipes were extracted by ultrasound assisted method,then Polygonatum filipes polysaccharides(CSPFPs)were obtained after elimination of the proteins with combined papain-Sevag method.The total sugar content was determined by phenol-sulfuric acid method.Structures of CSPFPs were analyzed by fourier transform infrared spectroscopy(FT-IR),high performance gel permeation chromatography(HPGPC)and high performance liquid chromatography(HPLC).Effects of CSPFPs on cell viability,pinocytic activity,TNF-α secretion,MAPK and NF-κB signaling pathways of RAW264.7 cells were explored by MTT,Neutral red,ELISA and Western blot,respectively.Results:Extraction rate of CSPFPs by ultrasound-assisted method was 41.61%,which contained total sugar content of 94.00%.CSPFPs with Mw of 3 125 Da was composed of arabinose(1.85%),galactose(6.14%),glucose(56.41%)and mannose(35.60%).The in vitro experiments showed that CSPFPs were non-cytotoxic and enhanced the pinocytic activity,TNF-α secretion and phosphorylation levels of p38,ERK,JNK,p65,IκB and IKK,indicating the activation of MAPK and NF-κB signaling pathways under the concentra-tion of 2.5～200 μg/ml.Conclusion:The ultrasound-assisted method can efficiently isolate CSPFPs with immunomodulatory activity,which provides basic data for the development and application of CSPFPs as an immunostimulant.

Objective:To separate and purify the polysaccharides from Polygonatum filipes,characterize their primary structure and investigate the immunomodulatory effects on RAW264.7 macrophages.Methods:Crude polysaccharides from Polygonatum filipes were extracted by ultrasound assisted method,then Polygonatum filipes polysaccharides(CSPFPs)were obtained after elimination of the proteins with combined papain-Sevag method.The total sugar content was determined by phenol-sulfuric acid method.Structures of CSPFPs were analyzed by fourier transform infrared spectroscopy(FT-IR),high performance gel permeation chromatography(HPGPC)and high performance liquid chromatography(HPLC).Effects of CSPFPs on cell viability,pinocytic activity,TNF-α secretion,MAPK and NF-κB signaling pathways of RAW264.7 cells were explored by MTT,Neutral red,ELISA and Western blot,respectively.Results:Extraction rate of CSPFPs by ultrasound-assisted method was 41.61%,which contained total sugar content of 94.00%.CSPFPs with Mw of 3 125 Da was composed of arabinose(1.85%),galactose(6.14%),glucose(56.41%)and mannose(35.60%).The in vitro experiments showed that CSPFPs were non-cytotoxic and enhanced the pinocytic activity,TNF-α secretion and phosphorylation levels of p38,ERK,JNK,p65,IκB and IKK,indicating the activation of MAPK and NF-κB signaling pathways under the concentra-tion of 2.5～200 μg/ml.Conclusion:The ultrasound-assisted method can efficiently isolate CSPFPs with immunomodulatory activity,which provides basic data for the development and application of CSPFPs as an immunostimulant.

OBJECTIVETo compare the efficacy differences between integrative therapy of electroacupuncture and repetitive transcranial magnetic stimulation (rTMS) and simple rTMS only in the treatment of comorbid anxiety and depression (CAD).

METHODSEighty-five cases of CAD were randomized into two groups. The observation group (40 cases) was treated with electroacupuncture at Baihui (GV 20), Yintang (EX-HN 3), Shenting (GV 24), Shenmen (HT 7) and Neiguan (PC 6) mainly, combined with rTMS. The control group (45 cases) was treated with simple rTMS. Separately, before treatment, after 5 days and 10 days treatment, Hemilton Depression Scale (HAMD) and Hemilton Anxiety Scale (HAMA) were used for the scoring of depression and anxiety. Additionally, the efficacies were compared between two groups.

RESULTSAfter 5 days, 10 days treatment, the scores of HAMA, HAMD were reduced remarkably as compared with those before treatment in two groups (all P < 0.001), HAMA score in observation group was lower than that in control group after 10 days treatment (P < 0.05). The remarkable effective rates of depression and anxiety in observation group those 10 days that treatment were 87. 1% (27/31) and 90.3% (28/31) respectively, which were higher apparently than 61.8% (21/34) and 61.8% (21/34) in control group separately (both P < 0.05).

CONCLUSIONThe integrative therapy of electroacupuncture and rTMS is definitely effective on CAD. This method is better than simple rTMS.

Adult ; Anxiety ; therapy ; Combined Modality Therapy ; Depression ; therapy ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Transcranial Magnetic Stimulation ; Treatment Outcome ; Young Adult

AIM: To investigate the role of brainstem auditory evoked potential(BAEP) in the early diagnosis of diabetic central neuropathy (DCN) and to probe into the relationship between plasma nitric oxide (NO) and DCN. METHODS: Experimental diabetes(ED) rat model was used. BAEP and plasma NO were measured dynamically after ED modeling. RESULTS: The peak latency (PL) of Ⅰ, Ⅲ, Ⅳ and V and the interpeak latency (IPL) of I-III and III-V of BAEP in ED group prolonged after 4 weeks of ED modeling (P＜0.05). All PL and IPL of BAEP prolonged after 6 weeks、8 weeks and 10 weeks (P＜0.01). The plasma NO increased to the top in ED group at 2 weeks and then decreased gradually. There was negative correlation between plasma NO and PL and IPL of BAEP. CONCLUSION: BAEP was sensitive and valuable in the early diagnosis of DCN. Reduction of plasma NO may play a role in the pathogenesis of DCN.

The abnormality of the central noradrenergic system in Alzheimer's disease has two cases: decrease or increase. The former is easy to be understood, because it is resulted from the outstanding forfeit of the noradrenergic neurons in the nucleus locus ceruleus (nLC); but the noradrenalin concentration does not change and enen increase, it seems to disagree with the loss of the noradrenergic neurons in the nLC. This article put emphasis on it, enumerated the related evidence and analyzed the possible causes. At present time, the drugs to increase noradrenalin is used mostly in clinic. This paper summarized it and put forth our own opinions.

AIM: To observe the relationship between noradrenaline and the synaptic configuration in rat hippocampus CA3 area. METHODS: Noradrenergic system injured models were produced by injecting 6-hydroxydopamine into rats' bilateral dorsal noradrenergic bundle. Then Y-type maze tests and elicitation of P3-like were carried out respectively before and after the model was established. The parameters of synaptic configuration in the hippocampus CA3 areas were also analyzed quantitatively. RESULTS: The decreases in the thickness of postsynaptic density material, the curvature of synaptic interface and the occurrence of perforated synapses, and the increase in the width of synaptic cleft were observed. CONCLUSION: The normal noradrenalin level in hippocampus is necessary to maintain the normal synaptic interface ultrastructure in hippocampus CA3 area.

AIM: To investigate the role of brainstem auditory evoked potential(BAEP) in the early diagnosis of diabetic central neuropathy (DCN) and to probe into the relationship between plasma nitric oxide (NO) and DCN. METHODS: Experimental diabetes(ED) rat model was used. BAEP and plasma NO were measured dynamically after ED modeling. RESULTS: The peak latency (PL) of Ⅰ, Ⅲ, Ⅳ and V and the interpeak latency (IPL) of I-III and III-V of BAEP in ED group prolonged after 4 weeks of ED modeling (P