Objective To explore the role of medial temporal lobe (MTL) structure in generating P 300.Methods Event-related potentials (ERPs) were performed in the patient following left lateral anterior temporal lobectomy and 17 health subjects. Two models of vision and auditus were involved in the study.Results The latency, amplitude and sideway distribution of auditory and visual P 300 were in the normal range gained from the healthy controls.Conclusion MTL structure may be not important in generating scalp P 300.

OBJECTIVE To investigate the antitumor effect of cadmium (Ⅱ) complex of pyrazolone derivatives 1-phenyl-3-methyl-4-propionyl-5-pyrazolone salicyloyl hydrazide-cadmium (Ⅱ) (Cd-PMPP-SAL) on the murine melanoma B16 cells in vitro and in vivo and its mechanisms. METHODS B16 cells were incubated with Cd-PMPP-SAL at 1.0, 1.5, 3.0, 5.0 and 10.0 mg·L-1 for 24, 48 or 72 h. The prolifera? tion rate of B16 cel s was evaluated by MTT assay. B16 cel s were incubated with Cd-PMPP-SAL at 6.25, 12.50 and 25.00 mg·L-1 for 24 h, while cell morphology was observed by Hoechst33258 staining. Apop?tosis of B16 cells was detected by Annexin Ⅴ-FITC/PI staining. The activity of caspases in B16 cells was detected by caspase activity assay. C57BL/6J mice were inoculated subcutaneously with B16 cells to establish a tumor-bearing model. Five days later, Cd-PMPP-SAL at 6.25, 12.50 and 25.00 mg·kg-1 was injected into tumors of C57BL/6J mice once a day for 12 d. The body mass was recorded daily. One day after the last administration, all the mice were killed and the tumor was harvested. Tumor volume and mass were measured, and the tumor inhibitory rates were calculated. Pathological changes of the tumor, liver and lung were observed under a microscope. The expressions of vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2) in tumor tissues were detected by immuno?histochemistry. The apoptotic cells in transplanted tumor tissues were detected by TUNEL. RESULTS Cd-PMPP-SAL inhibited the proliferation of B16 cells. The IC50 was 4.946 mg · L-1, and 95% confidence interval was 4.24-5.65 mg · L-1. The apoptosis rates(12.8 ± 1.4)% and (18.4 ± 0.4)% of Cd-PMPP-SAL 12.50 and 25.00 mg · L-1 groups were significantly higher than those of control group (1.7 ± 0.1)% (P<0.01). The activity of caspase 3 and 9 of Cd-PMPP-SAL 25.00 mg · L-1 group was significantly higher than that of control group (P<0.01), but there was no significant difference in caspases 3/7. The relative tumor volumes of Cd-PMPP-SAL 6.25, 12.50 and 25.00 mg · kg-1 treated groups from the 8th day of treatment were significantly decreased compared with the model group (P<0.01). The result of paraffin sections showed that the transplanted tumor tissues in Cd-PMPP-SAL 12.50 and 25.00 mg · kg- 1 groups exhibited different degrees of necrosis, but there was no significant pathological damage to the liver or lung tissues of mice. Compared with model group, expressions of VEGF and FGF2 in Cd-PMPP-SAL 12.50 and 25.00 mg · kg-1 treated groups were significantly inhibited (P<0.05), and apoptotic cell rates were significantly higher (P<0.05). CONCLUSION Cd-PMPP-SAL can inhibit growth of B16 cells in vivo and in vitro, which may be associated with induction of tumor cell apoptosis and inhibition of tumor angiogenesis.

Objective To study the effects of transcranial magnetic stimulation (TMS) on learning and memory, and angiogenesis and the dendritic structure of hippocampal CA3 pyramidal neurons after cerebral infarction. Methods Forty-eight male Sprague-Dawley rats were divided into a sham operated group, a model group and a TMS group (n = 16). Rat models of focal cerebral infarction were established with unilateral middle cerebral artery (MCA) suture occlusion in the model and TMS groups. The rats of the TMS group were given 4 weeks of TMS treatment beginning 1 day after the infarction (2 times per day, 30 pulses per time). Their learning and memory abilities were tested with a Y-maze. Angiogenesis and the dendritic structure of their hippocampal CA3 pyramidal neurons were detected after 4 weeks. Results Compared with the model group, learning and memory improved significantly in the TMS group. The average microvessel density of the hippocampus in the TMS group was significantly more than in the model group. The total length of apical dendrites of hippocampal CA3 pyramidal neurons in TMS group was significantly longer than in the model group. Conclusions The improved learning and memory observed following TMS treatment are likely to be related to changes in angiogenesis, the dendritic.structure of the hippocampal CA3 pyramidal neurons, and enhanced synaptic plasticity.

Objective To observe whether or not long-term inhalation of nitrous oxide can affect the cardiac function in rats. Methods Sprague-Dawley rats of either sex were randomly divided into four groups. Group A: inhaled common air for 50 days as control; Group B: inhaled 500mL/L N_2O for 15 days, two hours every day; Group C: inhaled 500mL/L N_2O for 30 days, two hours every day; Group D: inhaled 500mL/L N_2O for 50 days, two hours every day. Float glass intracellular microelectrode recording technique was used for observation of the duration (MAPD_(90), MAPD_(50) and MAPD_(20)) and amplitude of action potentials (APs) of left ventricular muscle cells in vitro. Angiotensin-2 (Ang-2) and eNOS were detected by immunohistochemical technique. Results ① Compared with that in Group A, the Aps duration of left ventricular muscle cells (MAPD) in Group B had no significant change while the MAPDs in Group C and Group D were extended significantly. There were no significant differences among the four groups in Aps amplitude. ② The expression of Ang-2 did not differ significantly between Group B and Group A. The expression level was higher in Group C and D than in Group A. ③ The expression level of eNOS was significantly lower in Group C and D than in Group A (P<0.05). Conclusion Long-term inhalation of N2O can significantly affect the cardiac function in rats, which may be related to higher expression of AngⅡin the heart induced by the long-time excitation of sympathetic nerves.

Objective To construct two DNA vaccines based on two glycoprotein antigen segments of Xinjiang hemorrhagic fever virus (XHFV) and to evaluate the immune responses in BALB/c mice following vaccination.Methods Two recombinant expression plasmids pVAX1-GcⅠand pVAX1-Gc Ⅱ were constructed by inserting XHFV YL04057 strain Gc Ⅰ (1 229-1 349 aa) and Gc Ⅱ (1 443-1 566 aa) fragments into the eukaryotic expression vector pVAX1 and then were identify by restriction enzyme digestion and sequencing analysis.The recombinant expression plasmids were transfected into mice by hydrodynamics-based transfection.Immune responses induced in mice were evaluated by testing the proliferation of T cells with MTT,measuring serum antibody level with ELISA and detecting cytokines in the supernatant of spleen cell culture with ELISA kit.Results The recombinant expression plasmids were successfully constructed as indicated by the results of restriction enzyme digestion and sequencing analysis.Expression of Gc Ⅰ and Gc Ⅱ genes in mice liver tissues was detected.Antibody titers in mice immunized with pVAX1-GcⅠor pVAX1-Gc Ⅱ were higher than those in mice immunized with pVAX1.Compared with pVAX1,pVAX1-Gc Ⅱ significantly enhanced the proliferation of splenic T lymphocytes and the expression of IFN-γ (P<0.01).Conclusion The constructed two DNA vaccines for XHFV can induce specific humoral and cellular immune responses in mice.pVAX1-Gc Ⅱ is better than pVAX1-GcⅠin immunogenicity and protective efficacy,suggesting that it can be used as a promising candidate for the development of DNA vaccine for XHFV.

The Crimean-congo hemorrhagic fever virus(CCHFV)is a geographically widespread fatal pathogen.Identification of the epitope regions of the virus is important for the diagnosis and epidemiological studies of CCHFV infections.In this study,expression vectors carrying series truncated fragments of the NP(nueleoeapsid protein)gene from the S fragment of CCHFV strain YL04057 were constructed.The recombinant proteins were expressed in E.coli and purified for detection.The antigenic of the truncated fragments of NP was detected with a polyclonai serum(rabbit)and 2 monoclonal(mAbs)(14B7 and 43E5)against CCHFV by Western-blot analyses.The results showed that the three expressed constructs,which all contained the region 235AA to 305AA could be detected by mAbs polyclonal serum.The results suggest that region 235-305 aa of NP is a highly antigenic region and is highly conserved in the NP protein.

OBJECTIVETo investigate the effect of chloroquine in reversing multidrug resistance (MDR) of HNE1/DDP cell line and explore the mechanism.

METHODSMTT assay was used to detect the cell viability of HNE1 and HNE1/DDP after exposure to different concentrations of DDP (2, 4, 8, 16 and 32 µmol/L) and different concentrations of chloroquine (5, 10, 20, 40, and 80 µmol/L). q-PCR was used to assess the expression of MDR1 mRNA and Western blotting was employed to detect P-glycoprotein (P-gp) expression in HNE1 and HNE1/DDP cells exposed to 5 and 10 µmol/L chloroquine. The cell apoptosis rate of HNE1 and HNE1/DDP cells exposed to 10 and 20 µmol

RESULTSChloroquine exposure caused dose-dependent suppression of the proliferation in both HNE1 and HNE1

CONCLUSIONChloroquine can reverse multidrug resistance in HNE1
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; Cell Line, Tumor ; Chloroquine ; pharmacology ; Down-Regulation ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans

The genomic DNA were extracted from the leaves of Polygonatum roseum (Liliaceae) in Xinjiang and the primers were designed according to conservative sequences of Polygonatum lectins gene. The complete ORF of Polygonatum roseum agglutinin (PRA) gene was amplified as a fragment of 550 bp, which was identical with predicted size. Like most of the plant lectin genes, there was no intron in the PRA gene. The ORF of the gene encoded 159 amino acid residues, in which included a signal sequence of 28 amino acid residues at its N-terminus. The cDNA sequence had 92% identities compared with the published sequence. The amino acid sequence and SWISS-MODEL analysis indicated that the three-dimensional structure of PRA strongly resembled with that of monocot mannose-binding lectins, which comprised with three antiparallel four-stranded beta-sheets arranged as a 12-stranded beta-barrel. The recombinant pGEX4T-1-PRA and pMAL-p2x-PRA prokaryotic expression vectors were constructed to produce GST-PRA and MBP-PRA fusion proteins in E. coli, respectively. SDS-PAGE of the fusion protein demonstrated that the PRA lectin protein migrated at a size of 14 kD. The immunization was performed by intra-muscular injection of pcDNA3-PRA, and the antiserum was detected by ELISA. Western blotting analysis showed the antiserum specifically bound the lectin protein. The establishment of such an expression system might provide materials for further investigation of the properties and functions of PRA proteins. It also laid the basis for plant genetic engineering on its defensive functions to pests and diseases.
Amino Acid Sequence ; Animals ; Base Sequence ; China ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Female ; Mannose-Binding Lectin ; biosynthesis ; genetics ; Mice ; Molecular Sequence Data ; Polygonatum ; chemistry ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Sequence Analysis

Objective To evaluate the clinical and pathological feature,as well as risk factors of lymph node metastasis (LNM) and high-volume LNM (hvLNM) in papillary thyroid microcarcinoma (PTMC) with di ameter ≤0.5 cm.Methods PTMC patients who received surgical treatments in Peking Union Medical College Hospital from Nov.2013 to Nov.2014 were reviewed.Patients were allocated into the ≤0.5 cm group and (0.5-1)cm group according to tumor diameter.Clinical and pathological features were assessed and compared.Risk factors of LNM and hvLNM were also assessed through univariate and multivariate analysis.Results 1414 patients were enrolled,of which 315 patients (22.3％) were in the ≤0.5 cm group.76 LNM (24.1％) and 9 hvLNM (2.9％) were detected in the ≤0.5 cm group.There was significantly less capsule invasion (14.3％ vs 25.0％,P＜0.05),LNM (24.1％ vs 39.8％,P＜0.05) and hvLNM (2.9％ vs 7.9％,P＜0.05) in ≤0.5 cm group than in (0.5-1)cm group.In univariate analysis,patients aging ＜40 years old were more likely to have LNM than those older than 40(38.0％ vs 20.1％,P＜0.05),while male patients tended to have more LNM than female (32.4％ vs 21.9％,P=0.073).No risk factors were identified for hvLNM.In multivariate analysis,multifocality and younger than 40 years old were the independent risk factors of LNM (OR=2.082 and 2.899,P＜0.05),while male tended to be the independent risk factors of LNM (OR=l.807,P=0.058).No independent risk factors was identified for hvLNM.Conclusions A certain proportion of PTMC patients are with tumor diameter ≤0.5 cm,who have lower risk of LNM and hvLNM.Dynamic observation may be an option,especially in older ≥40 years old),unifocal and female patients.

Objective To express and purify two domains GcⅠand GcⅡof Xinjiang hemorrhagic fever virus (XHFV) glycoprotein, and to study its immunogenicity and the effects on immune response in mice. Methods The prokaryotic expression plasmids of pET28a-GcⅠand pET32a-GcⅡwere constructed and transformed into E. coli BL21, respectively. The expression and purification conditions of rGcⅠand rGcⅡproteins were optimized. The antigenicity of the fusion protein was detected by Western Blot and enzyme-linked immunosorbent assay (ELISA). BALB/c mice were immunized by protein immunization and DNA priming-protein boosting. The mice were randomly divided into 5 groups, including pVAX1-GcⅠ+rGcⅠgroup, pVAX1-GcⅡ+rGcⅡgroup, rGcⅠgroup, rGcⅡgroup and saline group (control group) with 7 mice in each group. The serum antibody titer of mice was detected by indirect ELISA, and the immune effect was evaluated by spleen T lymphocyte proliferation assay and cytokine content determination. Results The fusion proteins rGcⅠand rGcⅡwere purified and obtained, which could react with positive serum of sheep and had good antigenicity. After three immunizations, the IgG levels in the serum of each experimental group were significantly higher than those in the control group (all P<0.001). The serum antibody titers of the experimental groups were reached above 1:12800. Among them, the concentration of Th2 type cytokine interleukin-4 (IL-4) in the spleen cell culture supernatant of rGcⅡ[(79.97±7.47) ng/L] and pVAX1-GcⅡ+rGcⅡgroup [(61.43±9.27) ng/L] was significantly higher than (24.29±3.81) ng/L of the control group, respectively (all P<0.01). The highest mass concentration [(42.46 ±2.60) ng/L] of Th1 type cytokine interferon-γ(IFN-γ) was observed in the pVAX1-GcⅡ+rGcⅡ group, which was significantly higher than (20.33±1.67) ng/L of the control group, and the difference was statistically significant (P<0.001). That showed a significant antigen-specific splenic T lymphocyte proliferation (P<0.001). Conclusions The purified recombinant proteins rGcⅠand rGcⅡhave good immunogenicity, which can make the immune system T lymphocytes tend to Th2 response, and pVAX1-GcⅡ combined with recombinant protein GcⅡ can induce better antigen-specific immune effect. And pVAX1-GcⅡ combined with recombinant protein GcⅡis expected to be used as vaccine candidates for the prevention and control of XHFV.