Objective To establish a new post-embedding immunoelectron microscopy method for localization of protein antigens in central nervous system using colloidal golds as probes.Methods Rat brain tissue sections were fixed with osmium-ferrocyanide mixture.Free floating frozen sections were collected to determine the retrievability of antigens by sodium metaperiodate(NaIO_(4)).Then the best concentration of NaIO_(4) for maximal antigen retrieval was determined on hydrophilic Technovit 7100 resin-embedded semithin sections.Finally,the corresponding antigen was labeled with colloidal golds on ultrathin sections.Results NaIO_(4) showed different retrieving effects on different antigens;to those antigens that could be effectively retrieved by NaIO_(4),low concentration of NaIO_(4)could realize antigen retrieval;EM microscopy showed that ultrathin sections treated by low concentration of NaIO_(4) allowed higher density of colloidal gold labeling without apparently compromising the ultrastructures of tissues.

Objective To study the expression pattern of RYK in the pathway of developmental corticospinal tract projection and provide hints for further study on RYK's function involved in this processes.Methods By In situ and immuno-fluorescence histochemistry,we examined the RYK expression pattern in the developmental murine motor cortex and the projection of corticospinal tract at various time points of E14.5、 E16、E18.5 and postnatal P3、 P5、P7.Results The motor cortex the RYK expression started at E18.5 stage and enhanced gradually at P0 stage.Moreover from P0 to P5 stage the RYK expression also appeared in the pathway of developmental corticospinal tract and decreased gradually after that.Conclusion RYK's expression is closely related to the corticospinal tract projection during its development stage.

Objective To study changes of expression in Ephrin and its receptors in DRG neurons after peripheral nerve injury. Methods A rat model of unilateral crushed sciatic nerve was prepared, and changes in the expression pattern of Ephrin B1, Eph B1、Eph B2、Eph B3、EphA4 and RYK in DRG neurons in the rats crushed unilateral sciatic nerve were estimated by the number of positive staining cells in DRG, and the percentage of positive cells as well as the cell size.Results After peripheral nerve injury, the expression of Ephrin B1 significantly decreased, but there was no change in expression of Eph B1、Eph B2、Eph B3 or Eph A4, RYK expression was intensely induced in DRG neurons. Conclusion Ephrin B1 and RYK showed very significant expression changes in ERG neurons after peripheral nerve injury, and may be a component factor process of peripheral never regeneration.

Objective:To study the effects of Yiqi Huoxue Ruanjian Jiedu(YHRJ) drug serum on Ca 2+ and mitochondrial membrane potential (△?m) in human hepatocellular carcinoma cell line BEL-7402.Methods: YHRJ decoction was composed of Astragalus Root, White Atractylodes Rhizome, Radix Notoginseng, Fructus Polygoni Orientalis, Radix Actindiae Argutae, Flos Campsis, Pangolin Scales, Oldenlandia diffusa Roxb, and Herba Scutellariae Barbatae. Twelve New Zealand rabbits were randomly assigned into 3 groups: NS group, YHRJ equivalent dose group(YHRJD) and YHRJ high dose group(YHRJG), which were fed with NS,YHRJ equivalent dose decoction (5 times as large as normal human dose) and high dose YHRJ decoction(10 times as large as normal human dosage), respectively. Drug serum was prepared after the rabbits were fed with drugs for 5 times. BEL-7402 cells were divided into 6 groups according to different drugs, including NS group, NS+DDP group, YHRJ equivalent dose group, YHRJ equivalent dose+DDP group, YHRJ high dose group, and YHRJ high dose +DDP group. The concentrations of NS,YHRJD and YHRJG were adjusted to 100 ml/L ,and the concentration of DDP was adjusted to 4.5 mg/L. Flow cytometry was applied to detect the concentrations of Ca 2+ and △?m 8, 12, 24 and 48 h after the drugs were added.Results: The Ca 2+ concentrations at 12, 24 and 48 h after treatment were significantly higher than those of the control group(P

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italic>Anoectochilus roxburghii (Wall.) Lindl. is a medicinal species belonging to the Orchidaceae, whose whole plant can be used as a medicinal herb, known as "JinXianLian". It has antidiabetic, liver-protecting, anti-inflammatory, etc. A. roxburghii has long been used as food and medicine in Guangdong and Fujian provinces. With the wide recognition of the concept of "medicine and food homology" and the surge of market demand, wild A. roxburghii has been far from meeting the supply. It is important to establish an artificial propagation system. Resource characteristics are the key basis for optimizing germplasm and propagation systems. Therefore, this paper summarizes the germplasm resource characteristics and propagation technologies of A. roxburghii in China to provide a reference for sustainable development and subsequent mechanistic research.

Objective To screen for the potential protein biomarkers in serum for the diagnosis of idiopathic Parkinson's disease (PD) using proteomic fingerprint technology. Methods Proteomic fingerprint technology combining weak cation exchange (WCX) magnetic beads with MALDI-TOF-MS was used to identify and compare the serum proteins from 61 patients with idiopathic PD, 29 patients with other neurodegenerative diseases (OND) and 30 healthy blood donors. Model of biomarkers and proteomics patterns associated with PD was analyzed by Biomarker Patterns Software. The model also was validated by 40 newly recruited PD cases. Results A total of 17 discriminating M/Z peaks which were related to PD were identified ( nonparametric test, Z:-4.039--2.633, P<0.01 ). Five biomarkers with M/Z of 6121, 5234, 2961,4309 and 8170 respectively generated an excellent model of distinguishing between PD and healthy groups. The sensitivity was 98.4% and the specificity was 83.1%. Blind testing in 40 newly recruited cases demonstrated a sensitivity of 85.0% (17 of 20 PD) and a specificity of 70. 0% (14 of 20 controls). Conclusions Combination of WCX magnetic beads with MALDI-TOF-MS is a useful method in establishing proteomic patterns associated with PD. It also may be used to construct a diagnostic model with PD Biomarkers. Although this model of biomarkers fails to distinguish between PD and OND controls, it is able to differentiate PD from healthy controls.

Based on ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), a rapid and simultaneous quantitative method for the measurement of seven components (kinsenoside; rutin; kaempferol-3-O-rutinoside; quercimeritrin; narcissin; isorhamnetin-3-O-glucoside; quercetin) of A. roxburghii was established. The separation was performed over 8.0 minutes on a Waters Acquity UPLC BEH Shield RP18 (2.1 mm × 100 mm; 1.7 μm) with a mobile phase consisting of acetonitrile (A) and 0.1% formic acid water solution (B) with gradient elution at a flow rate of 0.2 mL·min^-1; the column temperature was 30 ℃ and the injection volume was 2 μL. Electrospray spray ionization source (ESI source) was used for mass spectrometry, and positive and negative ion modes were detected at the same time. The results showed good linearity (R² ≥ 0.998 0), with good precision, repeatability and stability, and the average recovery was 97.71%-103.33%. Through cluster heat map and redundancy analysis, we found that kinsenoside was mainly distributed in stems, followed by leaves, and the lowest content in roots. The content of kinsenoside increased significantly in the stems of plants 6 months, but less change was evident in the roots and leaves. Flavonoids and flavonoid glycosides were mainly distributed in leaves. The UHPLC-MS/MS method established in this paper can be used for the quality control of A. roxburghii and provides a reference for establishing a more comprehensive quality detection method for this medicinal.

Objective To study the effects of repeated hypoxic exposures (HEs) on glycolysis, mitochondrial oxi-dative phosphorylation and energy charge in mouse brain. Methods Adult BALB/c mice were repeatedly exposed to hypoxia for 5 times and the standard tolerant time and body temperature were recorded. The activities of PFK, PK and mitochondrial complex Ⅰ in the brain were assayed. Phosphoadenosines and energy charge were measured. Results Repeated HEs prolonged the hypoxic tolerance and reduced the body temperature. The activities of PFK and PK experienced regular changes, with an increase in 1st and 3rd HEs and a decline to control levels in 5th HE. The complex Ⅰ activity continued to decrease during HEs. The energy charge was stable. Conclusion HEs lead to a regular change of glycolysis, a continued inhibition of mitochondrial oxidative phosphorylation, and a main-tained energy charge in the brains of mouse.

Objective Toanalyzethealterationsofplasmaglucocerebrosidase(GBA),protein phosphatase 2A (PP2A)and its degradation product ceramide in patients with ischemic stroke. Methods Atotalof45inpatientswithischemicstrokeattheDepartmentofNeurology,theAffiliated Hospital of Logistics College of the CAPF were enrolled from May to September 2013,and 45 age-and sex-matched healthy subjects at the Physical Examination Center collected at the same time period were used as a control group. Blood samples of the patients and healthy subjects were obtained,anticoagulated, and the plasma was separated. H50 protein chip and laser matrix-assisted laser desorption/ionization top of flymassspectrometrywereusedtotestthelevelsofplasmaceramide.Results TheplasmaGBAand PP2A activities in patients with ischemic stroke were significantly lower than those of the control group;the GBA activities of the ischemic stroke group and the control group were 2 . 4 ± 0 . 8 and 3 . 1 ± 1 . 4 U/L respectively. There was significant difference (P<0. 05);the PP2A activities of the two groups were 6. 5 ± 2. 8 and 14. 5 ± 4. 7 U/L respectively (P<0. 01). The relative level of the plasma ceramide in patients with ischemic stroke was 1. 9 ± 0. 7,and it was significantly lower than 12. 2 ± 5. 0 of the control group (P<0.01).Conclusion ThedecreasedlevelsofplasmaGBAandPP2Aactivitiesaswellasthe ceramide in patients with ischemic stroke suggested that the abnormal phosphorylation of synuclein in the blood of patients with stroke.