Objective To study the effects of Multi-Walled Carbon Nanotube with different doses on the cellular proliferation of human embryo kidney 293 cells.Methods The cultured human embryo kidney cell line 293 was seeded into 96-well plates and various concentrations of Multi-Walled Carbon Nanotube were added into different groups of culture.Methyl thiazolyl tetrazolium(MTT)assay were applied to detect the cellular viabilities after being incubated with Multi-Walled Carbon Nanotube for 48 hours and the results were calculated with specific static software to protract the cell viability curve.Results The Multi-Walled Carbon Nanotube had different effects of inhibiting the multiplies of 293 cells depending on its concentrations.In addition,when the concentration of Multi-Walled Carbon Nanotube reached over 0.5ug/ml,the inhibition became significantly.Conclusions Multi-Walled Carbon Nanotube can penetrate the cell membrane of 293 cells to influence the activities of cells and the ability of proliferation will be decreased significantly when the concentration came to some degree.Therefore,the safety dosage of the Multi-Walled Carbon Nanotube on human normal embryo kidney 293 cells were estimated in this research.

The projections from the nucleus tractus solitarii (NTS) and the nuclei parabrachiales (PB) to the nucleus accumbens (Acc) in the rat have been visulized using anterograde and retrograde HRP tracing techniques. After injecting HRP into the Acc, retrogradely labelled neurons were observed in bilateral PB and NTS with ipsilateral predominance. The labelled neurons were concentrated in the following areas of the PB and NTS: the waist area of the caudal PB, the external subnucleus and other part of the nucleus medialis parabrachialis (PBm), the external, central, and internal subnuclei of the nucleus lateralis parabrachialis (PB1) and the medial subnucleus of the caudal NTS (NTSm). After injecting HRP into the NTSm and commissural nucleus of the NTS, anterogradely labelled terminals were found bilaterally in the PBm and the ventral 3/4 area of the PB1. The densest sites occupied the waist area, the external, central, and internal subnuclei of the PB1. The density of the labelled terminals on the ipsilateral side was little higher than that on the contralateral side. The results indicate that there are two possible pathways from the NTS to the Acc, the one is the direct projection from the medial subnucleus of the caudal NTS to the Ace, the other is an indirect one, i. e. from the medial subnucleus and commissural nucleus of the caudal NTS to the waist area of the caudal PB, the external subnucleus, the dorsal part of the PBm, the external, central, and internal subnuclei of the PB1, where the NTS projection is. presumed to be relayed, and then, project from the PB to the Acc. This connection may be involved in the neural regulation of visceral and locomotor activities.

Objective:To sduty the effect of β-lactam antibiotics efiriaxone on the levels of glutamate and glutamate transporter subtype-1 in hippocampus following traumatic brain injury in rats.Methods: All rats were divided randomly into three groups:sham group;TBI group and CTX group.The rat model of TBI were made by modified Feeney method ,and treated with ceftriaxone immediately after injury ( 200 mg/kg ).Wet-dry weight method was used to evaluate brain edema.The content of glutamate in hippocampus was measured with the high performance liquid chromatography.The expression of GLT-1 in hippocampus areas was tested by immunohisto-chemical and Western blot methods.Results:Compared with TBI group ,TBI-induced cerebral edema was significantly attenuated ( P<0.05 ) , the content of glutamate in hippocampus was were significantly decreased ( P<0.05 ) , the level of GLT-1 were significantly increased in CTX group ( P<0.05 ).Conclusion: β-lactam antibiotics ceftriaxone can block the excitatory neurotoxicity , reduce the extent of brain edema.

ObjectiveTo investigate the effects of HO-1 on mast cells degranulation during liver ischemia/reperfusion injury.MethodsSD rats ( n =20) were randomly divided into 4 groups : ( 1 ) sham operation group, (2) ischemia/reperfusion injury group (I/RI group), (3) CoPP group, CoPP was given before 24 h(5 mg/kg), (4) ZnPP group, ZnPP was given before 24 h (20 mg/kg). The rat model of liver ischemia/reperfusion was built. Samples were collected after 2 hours reperfusion. HO-1 mRNA expression was assessed by RT-PCR. The expression of HO-1 protein was assessed by Western blot. Serum ALT, AST levels were determined. The amount of degranulated mast cells were detected by toluidine blue staining. Liver injuries were evaluated by HE staining.ResultsCompared to sham group, liver HO-1mRNA and protein expression, ALT, AST levels, the number of degranulated mast cells significantly increased in I/RI group, CoPP group and ZnPP group. Moreover, liver injuries were more serious. Compared to I/RI group, there were increased HO-1mRNA and protein expression, reduced ALT, AST levels and the number of degranulated mast cells, also reduced liver injuries were found in CoPP group. Compared to I/RI group, the expression of HO-ImRNA and protein was down-regulated, ALT, AST levels elevated and the number of degranulatedmastcellsincreasedaccompanyingmoreseriousliverinjuriesinZnPPgroup.ConclusionsHO-1 overexpression reduces liver ischemia/reperfusion injury, possibly by inhibiting mast cells degranulation in liver tissues.

The expression of paired immunoglobulin-like receptor B (PirB) in normal and injured spinal cord of rats was investigated. The SD rat hemi-sectioned spinal cord injury (SCI) model was established. Before and 1, 3, 7, 10 days after SCI, the spinal cord tissues were harvested, and Western blot and immunohistochemistry were used to examine the expression and location of PirB. The results showed that the expression level of PirB in the normal spinal cord of SD rats was low. At the first day after SCI, the expression of PirB was obviously increased, and that in the injured spinal cord from the first day to the 10th day was significantly higher than in the normal spinal cord. The positive expression of PirB in neurons from different regions of gray matter of the injured spinal cord was seen. It was concluded that the expression of PirB in the normal spinal cord of rats was low. The expression of PirB in SCI was significantly increased till at least the 10th day.

To construct a lentiviral shRNA vector targeting human protein phosphatase 1D magnesium-dependent (PPM1D) gene and detect its effectiveness of gene silencing in human gliomas, specific siRNA targets with short hairpin frame were designed and synthesized. DNA oligo was cloned into the pFU-GW-iRNA lentiviral expression vector, and then PCR and sequencing analyses were conducted to verify the constructs. After the verified plasmids were transfected into 293T cells, the lentivirus was produced and the titer of virus was determined. Real-time quantitative PCR and Western blot were performed to detect the PPM1D expression level in the infected glioma cells. PCR and Western blot analyses revealed the optimal interfering target, and the virus with a titer of 6×10(8) TU/mL was successfully packaged. The PPM1D expression in human glioma cells was knocked down at both mRNA and protein levels by virus infection. The expression of PPM1D mRNA and protein was decreased by 76.3% and 87.0% respectively as compared with control group. The multiple functions of human glioma cells after PPM1D RNA interference were detected by flow cytometry and cell counting kit-8 (CCK-8). Efficient down-regulation of PPM1D resulted in significantly increased cell apoptosis and reduced cell proliferation and invasion potential in U87-MG cells. We have successfully constructed the lentiviral shRNA expression vector capable of stable PPM1D gene silencing at both mRNA and protein levels in glioma cells. And our data gave evidence that the reduced cell growth observed after PPM1D silencing in glioma cells was at least partly due to increased apoptotic cell death.

Objective To explore the viral load levels after HIV/AIDS merges HBV/HCV infection and its relationship with T lymphocytes .Methods HIV RNA ,HBV DNA ,HCV RNA ,CD4+ T lymphocyte frequency ,CD8+ T lymphocyte frequency ,CD4/CD8 measured in HIV/AIDS simple infection group ,mixed HIV/HBV infection group and mixed HIV/HCV infection group .Ana‐lyze relationship of T lymphocyte and HIV RNA ,the correlation of HBV DNA/HCV RNA ,HIV RNA ,CD4+ T lymphocyte fre‐quency ,CD8+ T lymphocyte frequency ,CD4/CD8 .Results CD4+ T lymphocyte frequency of HIV/AIDS simple infection group , mixed HIV/HBV infection group and mixed HIV/HCV infection group showed negative correlated with their respective group′s HIV RNA(P<0 .05);CD8+ T lymphocyte frequency of HIV/AIDS simple infection group and mixed HIV/HBV infection group showed negative correlated with their respective group′s HIV RNA(P<0 .05);CD4/CD8 of HIV/AIDS simple infection group , mixed HIV/HBV infection group and mixed HIV/HCV infection group show negative correlated with their respective group′s HIV RNA(P<0 .05) .Conclusion T lymphocyte of HIV/AIDS patients drop faster ,leading to high viral load of HIV RNA ,HBV DNA and accelerating HIV disease progress with HBV infection .T lymphocyte of HIV/AIDS patients with HBV infection drop faster than with HCV infection .

Stribild is a compound preparation composed of elvitegravir, cobicistat, emtricitabine and tenofovir. As for the first-treated AIDs patients or re-treated patients with creatinine clearance≥70ml·min-1 , it has definite clinical effect. Owing to once a day, stribild can enhance the acceptability of the patients, and as for the patients treated with other regimen complicated with diarrhea, pain in stomach and numbness in hands and feet, it also provides a good option. However, its adverse effects on kidney, liver and bone density in the long term use should be paid attention.

Objective To analyze the differences of immune responses against Mycobacterium tu-berculosis antigens induced in two different nonhuman primates and to provide rationales for the selection of suitable animal models for vaccine efficacy evaluation.Methods Expression of functional surface markers including CD69 and HLA-DR, the activation markers on CD4+and CD8+T cells from in rhesus macaques and cynomolgus monkeys were measured by flow cytometry analysis.PBMCs were isolated from rhesus ma-caques and cynomolgus monkeys with Mycobacterium tuberculosis infection and stimulated with PPD and pep-tide pools ( ESAT-6/CFP-10) .Enzyme-linked imunospot ( ELISPOT) assay was performed to detect IFN-γproducing lymphocytes.Results The CD4+and CD8+T cells isolated from rhesus macaques without Myco-bacterium tuberculosis infection expressed higher levels of CD69 and HLA-DR than those from healthy cyno-molgus monkeys (P<0.01).The numbers of IFN-γspot forming cells/106 PBMCs in rhesus macaques with Mycobacterium tuberculosis infection for 10 and 11 months were respectively 3 and 3.5 times higher than that of cynomolgus monkeys upon after the stimulation of PBMCs with PPD.The levels of IFN-γproduction by the cells from rhesus macaque group were also higher than those from cynomolgus monkey group upon after the stimulation of PBMCs with ESAT-6 or CFP-10 peptide pools.Conclusion More IFN-γproducing cells were induced in rhesus macaques than that in cynimolgus monkeys after stimulation with Mycobacterium tu-berculosis antigens.Therefore, the rhesus macaques might be a better animal model for evaluating immune responses induced by Mycobacterium tuberculosis vaccines.

Designable experiment refers to such an experiment that students apply their knowledge and experiment skills to design and prepare experimental materials,instruments,means and procedures based on specific purpose and requirements,and finish the formal experiment by themselves.Through designable experiment,students' integration capability such as applying knowledge,manual operation and handling problem are cultured.We explore setting up designable experiment in the course of clinical hematological examination,and evaluate the effectiveness of teaching.