Objective To fractionate phosphoproteome of mouse liver by two-dimensional (2D) liquid phase chromatography fractionation. Methods Phosphoproteins were extracted from lysates of normal mice livers by phosphate metal affinity chromatography (PMAC) resin. The phosphoproteins were exchanged by start buffer and separated by chromatofocusing in the first dimension. Then the fractions between pH 8.5 and pH 4.0 were separated by non-porous silica (NPS) reverse-phase high performance liquid chromatography (RP-HPLC). Finally, the UV maps were converted into gel-like maps by ProteoVue software. Results Phosphoproteins of mouse liver were successfully extracted and fractionated by two dimensional liquid phase chromatographic fractionation after concentration and desalt. Then pI/UV map of mouse liver phosphoproteome was successfully set-up. There are 16 fractions between pH 8.5 and pH 4.0 after chromatofocusing in the first dimension and the UV maps of each fraction were converted into pI/UV gel-like maps. Conclusions Combination of technique of phosphoproteins enrichment and 2-D liquid phase chromatographic fractionation is an effective approach to research phosphoproteome and the key base for further identification and investigation of phosphoproteins.

Aim To explore the effects of acupuncture and sinomenine on the withdrawal symptoms,on the spontaneous discharge and synaptophysin expression of amygdala of heroin-addicted rats.Methods 75 SD rats were randomly and equally divided into 5 groups consisted of control,heroin-addicted,acupuncture,sinomenine and combined treatment group.The model of heroin-addicted rats were established with heroin subcutaneous injection,which in acupuncture,sinomenine and combined group were subsequently punctured at 3 acupoint(Neiguan,Baihui and Zusanli),intramuscular injected with sinomenine and treated with combined methods(acupuncture and sinomenine)respectively.The withdrawal symptoms of rats in different group were evaluated,and then the spontaneous discharges and synaptophysin expression of amygdala were detected with electrophysiological and immunohistochemical technology respectively.Results The heroin-addicted rats were successfully established,being suggested by typical withdrawal symptoms induced by naloxone(P<0.01 vs control),which were alleviated in the 3 treatment groups(acupuncture,sinomenine and combined ) with varying degree(P<0.01 vs heroin group).The electrophysiological results suggest that the forms of low-frequency and single irregular discharges in amygdala were increase,bunchy/cluster and complex discharges were decreased(P<0.05 vs control),which were reversed apparently in sinomenine and combined treatment groups(P<0.05 vs heroin group),closed to control.The immunohistochemical results show that expression of synaptophysin in amygdala was increased obviously(P<0.01 vs control),which was decreased in the 3 treatment groups(acupuncture,sinomenine and combined),P<0.01 vs addicted group.Conclusion Treatments of acupuncture and sinomenine administration can alleviate withdrawal symptoms of heroin-addicted,and rectify the abnormal spontaneous discharges and expression of synaptophysin in amygdala of heroin-addicted rats for certain degree,especially with combined treatments.

Objective:To investigate the effect of the new nanomaterials,polyamidoamine dendrimers PAMAM-D,mediated foreign gene human decay accelerating factor(hDAF) for xenotransplantation transfected pig sperm cells.Methods:PAMAM-D/hDAF cDNA compounds were made.The compounds were divided into 0.2 μg,0.4μg,0.6μg,0.8 μg and 1.0μg groups (each group adding corresponding dose of PAMAM-D in accordance with the N/P ratio 10:1,20:1,40:1),then were digested by restriction enzymes.The compounds were incubated with washed pig sperm cells.Then the transfection efficiency was detected by in situ hybridization in the different groups.Results:The PAMAM-D molecule can prevent electrophoretic migration of DNA in the compound.After digested the compounds by restriction enzymes,DNA can not be degraded.The transfection efficiency was different in different groups.Among the total,the efficiency was higher in both groups of 0.4 μg and 0.6μg than that of others.The top was the group of 0.4 μg linear plasmid plus PAMAM-D when the N/P ratio was 20:1(47.5%±O.2%,167% vs control group).Conclusion:PAMAM-D can improve the efficiency of exogenous gene transfeeted pig sperm cells,which can reinforce the stable binding of exogenous DNA to sperm cells.

Objective: To analyze the magnetic resonance imaging (MRI) radiomic performance of hypoxic area in nasopharyngeal carcinoma patients, aiming to provide a reference for identification and analysis of hypoxic area. Methods: The MRI-T₁, MRI-T₂, MRI-T₁+ and PET/CT images of 32 patients initially diagnosed with nasopharyngeal carcinoma were retrospectively analyzed. The gross tumor volume (GTV) of nasopharynx was countoured and the hypoxic areas (GTV-H) were identified by ¹⁸F-FMISO-PET images. The non-hypoxic areas (GTV-NH) were defined as the rest of areas removed GTV-H from GTV. The radiomic features of GTV-H and GTV-NH were extracted and compared. Results: The average volume of GTV-H and GTV-NH was (10.92±11.02) cm³ and (7.21±5.70) cm³, respectively. The maximum rate of change was 46% for intensity direct-global min (ID-GM) on MRI-T₁(P<0.05, AUC>0.7 and Youden index>0.5). The average rate of change was 136% for long run emphasis (LRE), long run high gray level emphasis (LRHGLE) and long run low gray level emphasis (LRLGLE) on MRI-T₂(P<0.05, AUC>0.7 and Youden index>0.5). The high change rates was greater than 90% on MRI-T₁+ (P<0.05, AUC>0.7 and Youden index>0.5) for ID-GM, LRE, LRHGLE and LRLGLE. Conclusions The hypoxic area of tumor target can be reflected by MRI radiomics on T₁/T₂/T₁+ . Quantifying and tracking the variations of these features can bring benefit to recognize the hypoxic area of nasopharyngeal carcinoma tumor target.

OBJECTIVE：To study the effects and potential mechani sm of deoxyschizandrin on the proliferation ，migration and invasion of nasopharyngeal carcinoma cell HONE- 1. METHODS ：HONE-1 cell was set as cell model ，while CCK- 8 test，wound healing assay and Transwell chamber test were used to detect the proliferation ，migration and invasion ability changes of HONE- 1 cells after treatment with different concentrations [ 0（blank control ），10，20，40 μmol/L] of deoxyschizandrin. Computer molecular docking was performed to analyze the binding ability between deoxyschizandrin and Met protein. Western blotting assay was used to detect the relative protein expressions of p-Met ，p-PI3K，p-Akt，Bcl-2 and N-cadherin in cells. RESULTS ：Compared with blank control ，the proliferation ，migration and invasion ability of cells after treated with 10，20，40 μmol/L deoxyschizandrin were all decreased significantly （P＜0.05）. Results of molecular docking revealed that deoxyschizandrin could stably bind with the activity pocket of Met protein. Results of Western blotting assay demonstrated that compared with blank control ，10，20，40 μmol/L deoxyschizandrin all decreased the relative protein expressions of p-Met ，p-PI3K，p-Akt，Bcl-2 and N-cadherin in cells significantly（P＜0.05）. CONCLUSIONS ：Deoxyschizandrin can inhibit the proliferation ，migration and invasion of HONE- 1 cell via inhibiting the activation of Met/PI 3K/Akt signaling pathway.