Female ; Humans ; Methotrexate ; Pregnancy ; Pregnancy, Ectopic

Objective To investigate the effects of different abstinent duration on brain functional network of heroin addicts at resting state.Methods Sixteen heroin addicts during protracted abstinence for 11-13 months (PA12 group) and twenty heroin addicts during protracted abstinence for 5-7 months (PA6 group) were recruited in the resting-state functional MRI study.Graph theoretical methods were applied to construct topological organization of whole brain network and nodes betweenness of the networks in all subjects,and the between-group differences were analyzed.The correlation of the node betweenness with the abstinence duration was conducted.Results There was no significant difference (P＞0.05) in the small world characteristic (γ≈1,λ》1) between two groups.Compared with the PA6 group,the PA12 group demonstrated significantly decreased nodal betweenness in regions of left parahippocampal gyrus,left precentral gyrus,and significantly enhanced nodal betweenness in regions of the left cuneus,left temporal and right middle occipital gyrus (all P＜0.05).Moreover,the betweenness of the left precentral gyrus (r =-0.52,P =0.001) and parahippocampal gyrus (r=-0.49,P=0.002) were negatively related with the abstinence duration,the betweenness of the right middle occipital was positively correlated with the abstinence duration (r=0.49,P=0.003).Conclusion The brain network small world topology of heroin addicts tend to be stable after 5-7 months of abstinence.Long-term abstinence may minimize the addicfs memory of drugs and potential drug seeking behavior,and recover addicts' visual spatial attention function.

Objective To examine the cognitive function in healthy first-degree relatives (FDRs) of patients with bipolar Ⅰ disorder.Methods Cognitive function were studied in one hundred twenty healthy FDRs of patients with bipolar Ⅰ disorder and one hundred normal controls using digital symbol, digital span, visual reproduction, trail making test A (TMT-A) and trail making test B (TMT-B).Results Compared with normal controls, FDRs showed impairment in all indexes of the tests, including digital symbol, digital span (forward, reversed and forward + reversed), visual reproduction, TMT-A and TMT-B (t=-3.44、-4.23、-4.32、-4.98、-2.59、4.32、3.78, respectively, Ps≤0.01).By analysis of covariance (covariant: age and years of education), FDRs were still impaired in these indexes (P<0.05).Compared with sex-matched normal controls, male FDRs showed impairment in all indexes of the tests, but female FDRs only showed impairment in digital span (reversed, forward + reversed), TMT-A and TMT-B (P<0.05).Conclusions Attention, memory and executive function are impaired in healthy first-degree relatives of patients with bipolar Ⅰ disorder.

Objective To investigate the effect of Pingfei Oral Liquid (POL) on the distribution of mast cells (MCs) and the expression of interleukin 6 (IL-6) in the lung tissue of radiation pneumonia rats. MethodsForty-five SD rats were randomized into 3 groups : normal control,model group and POL group. The rat model of radiation lung fibro-sis was set up by a single X-ray dose of 20Gy irradiation over the whole chest of the rats. POL (20 g·kg-1·d-1,once a day, five times a week) was given orally one week before irradiation and the treatment lasted 5 weeks. MCs in the lung tissue were stained with toluidine blue firstly and then were counted 2, 4 and 8 weeks after irradiation. IL-6 protein expression of lung tissue was measured by immunohistochemical assay 8 weeks after irradiation, and mRNA ex-pression was determined with RT-PCR 4 weeks after irradiation. ResultsIt's showed the aggregation of large amount of pulmonary mast cells and increase of IL-6 protein expression 8 weeks after irradiation (P < 0.01).IL-6 mRNA expression in the irradiated lung of rats increased 4 weeks after irradiation (P < 0. 01). POL could reduce the aggrega- tion of MCs (P < 0. 01) and the expression of IL-6 protein (P < 0. 01) and mRNA (P < 0. 05) in the lung tissue. ConclusionPOL can prevent radiation pneumonia in rats by reducing the aggregation of mast cells and inhibiting IL-6 expression in the lung tissue.

Objective To evaluate the relationship between newly developed depression after mild traumatic brain injury (mild-TBI)and abnormality on susceptibility-weighted imaging (SWI).Methods One hundred and eighty patients after mild-TBI,who had normal findings at conventional CT and MRI were enrolled in our study.All cases underwent conventional MRI and SWI.Ac-cording to SWI,the number and volume of microbleed lesions were calculated.All patients were followed up within 1 year after TBI,and divided into depressive group and non-depressive group based on SCID-IV.The difference in microbleed lesions on SWI was compared between the depressive group and non-depressive group.Results The incidence rate of microbleed on SWI in depres-sive group was higher than that in non-depressive group (P < 0.001 ).Among patients with microbleed lesions,the volume and number of lesions in depressive group were more than those in non-depressive group (P <0.001),and these differences were only found at the frontal,parietal and temporal lobes (P < 0.001 ).No significantly difference between depressive and non-depressive group was found in other areas (P >0.05).Conclusion SWI is useful to identify the microbleed lesions after mild TBI.The depres-sion after TBI is correlated with the location and distribution range of microbleed lesions.

To investigate the effects and potential mechanisms of hyperoside (Hyp) on the vascular endothelium function in middle cerebral artery (MCA) ex vivo in rats. Isolated arterial segments from MCAs of rats were used for surveying vasomotoricity in a pressurized chamber. Transmembrane potential was recorded by using glass microelectrodes to evaluate hyperpolarization. Hyp (1 x 10(-6)-1 x 10(-4) mol . L-1) was utilized to observe the effect on 1 x 10(-7) mol . L-1 U46619-preconstricted MCA in rats. The results showed that 1 x 10(-6)-1 x 10(-4) mol . L-1 Hyp significantly induced concentration-dependent vasodilatation and hyperpolarization, leading to the maximal diastolic ratio of (73. 2 ± 6. 1)% and maximal changes in membrane potentials of (-13. 2 ± 2. 2) mV. Hyp still elicited vasorelaxation and hyperpolarization by removal of endothelium in MCA of rat, which was notably attenuated as compared with vascular endothelium-intact group (P <0. 01). In the MCAs preconstricted by U46619 (1 x 10(-7) mol . L-1), Hyp (1 x 10(-6)-1 x 10(-4) mol . L-1) produced concentration-dependent vasorelaxation and hyperpolarizition that were partially attenuated by 3 x 10(-5) mol . L-1 L-NAME(a NOS inhibitor) plus 1 x 10(-5) mol . L-1 PGI2 ,(a synthetase inhibitor). The residual effects were further decreased by 1 x 10(-3) mol . L-1 TEA (an inhibitor of Ca2+-activated potassium channel) or 1 x 10(-5) mol . L-1 PPG (a blocker of endogenous H2S synthese-CSE). Similarly, 1 x 10(-5)-1 x 10(-3) mol . L-1 NaHS (a donor of exogenous H2S) or 1 x 10(-5)-1 x 10(-3) mol . L-1 L-Cys (the substrate of endogenous H2S synthesis) obviously evoked dose-dependent vasodilatation and hyperpolarization of MCA in rats. These findings indicated that Hyp may induce endothelium-dependent and endothelium-independent responses. And the endothelium-dependent vasodilatation may be related to the increases of endogenous H2S that has been promoted Hyp in the endotheliocyte of MCAs, and activated Kca and opening of Kca channels, resulting in the hyperpolarization of vascular smooth muscle cell membrane and subsequent reduction of Ca2+ influx and vasodilation.
Animals ; Endothelium, Vascular ; drug effects ; Enzyme Inhibitors ; pharmacology ; Female ; Male ; Middle Cerebral Artery ; Muscle, Smooth, Vascular ; drug effects ; Nitric Oxide ; metabolism ; Potassium Channels ; metabolism ; Quercetin ; analogs & derivatives ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfides ; metabolism ; Vasodilation ; drug effects ; Vasodilator Agents ; pharmacology

Anesthetics, Intravenous ; administration & dosage ; pharmacology ; Animals ; Animals, Newborn ; Cerebral Cortex ; cytology ; metabolism ; Drug Synergism ; Female ; Fentanyl ; administration & dosage ; pharmacology ; Male ; Midazolam ; administration & dosage ; pharmacology ; Neurons ; metabolism ; Patch-Clamp Techniques ; Primary Cell Culture ; Rats ; Rats, Sprague-Dawley ; Voltage-Gated Sodium Channels ; drug effects

Objective To evaluate the influence of ultrasound-mediated contrast agent microbubbles carrying kallidinogenase targeted therapy on neurogenesis and angiogenesis after experimental acute cerebral infarction.Methods Kallidinogenase-loaded microbubbles were prepared using mechanical shaking method.Middle cerebral artery occlusion (MCAO) model was established in male Wistar rats by sutureoccluded method.MCAO rats (n =120) were randomly divided into 5 groups:ultrasound-mediated kallidinogenase-loaded microbubbles group (group 1),ultrasound-mediated microbubbles group (group 2),ultrasound-mediated saline group (group 3),kallidinogenase group (group 4),saline group (group 5).Medication was given through tail vein daily for 2-6 consecutive days starting 24 h after MCAO.Ultrasound groups (groups 1,2 and 3) were given 2 MHz pulse ultrasonic irradiation on the ischemia lateral skull for 10 min after injection.Cell proliferation was examined using 5'-bromo-2'-deoxyuridine (BrdU,50 mg/kg).Infarction volume and neurological function were evaluated on the 3rd,7th day after MCAO respectively.Doublecortin (DCX) + cells in the subventricular zone and laminin + in the peri-infarction region were observed at the same time.Results The number of DCX + cells in the subventricular zone (under 20 × ocular) of groups 1-5 was 251.8 ± 13.1,125.7 ± 11.6,130.2 ± 13.7,234.5 ± 12.4 and 123.7 ± 10.0 respectively.The percentage of laminin + cells in the peri-infarction region (× 10 objective) of groups 1-5 was 10.0％ ± 0.8％,5.2％ ± 0.7％,5.0％ ± 1.0％,8.0％ ± 1.8％ and 5.0％ ± 0.9％ respectively.The number of DCX + cells in the subventricular zone and laminin + cells in the peri-infarction region of the group 1 and the group 4 was significantly more on the 7th day after MCAO compared with those of the other three groups (DCX:t values of group 1 vs groups 2,3,5 were 17.88,17.17 and 18.16,all P ＜ 0.01 ; t values of group 4 vs groups 2,3,5 were 15.42,14.78 and 15.70,all P ＜0.01.Laminin:t values of group 1 vs groups 2,3,5 were 7.01,6.71 and 7.11,all P ＜ 0.01 ; t values of group 4 vs groups 2,3,5 were 4.23,3.94 and 4.33,all P ＜0.01).Moreover,the number of DCX+ cells in the subventricular zone (t =2.46,P ＜ 0.05) and laminin + cells in the peri-infarction region (t =2.78,P ＜ 0.05) of the group 1 was much more than those of the group 4.Neurologic scores on the 7th day of groups 1-5 were 1.00 (0.75,1.25),2.0 (2.00,3.00),2.0 (1.00,2.00),1.5 (0.75,2.00),2.0 (2.00,2.50).Compared with other four groups,group 1 showed better functional improvement after stroke (U values of group 1 vs groups 2-5 were 2.0,4.0,7.5 and 2.5,all P ＜ 0.05).While there was no significant difference in infarction volume among five groups at all the time points after MCAO.Conclusions Our study demonstrates ultrasound-mediated kallidinogenase-loaded contrast agent microbubbles targeted therapy promotes neuroblasts proliferation and vascular regeneration compared with mediated and non-medicine-loaded microbubbles therapy,which attributes to functional improvement after MCAO.Therefore,ultrasound-mediated ultrasound contrast agent microbubbles carrying drugs targeted therapy may have a perspective on ischemic stroke.

BACKGROUND:Stable and efficient labeling of dental pulp stem cel s in vitro is most important in tracer technique, which is also the basis of tooth regeneration in vivo.
OBJECTIVE:To determine the optimal condition and method for transfection of stem cel s derived from rat dental pulp with green fluorescent protein infection by lentiviral vector and to determine whether green fluorescent protein-labeled dental pulp stem cel s maintain their stem cel properties.
METHODS:Rat dental pulp stem cel s were obtained by modified enzyme digestion method, to identify the immune phenotype and differentiation potential. Dental pulp stem cel s were infected with green fluorescent protein by lentiviral vector for 24 and 48 hours at different multiplicity of infection (MOI) (5, 10, 25, 50 and 100). The infection efficiency and fluorescence intensity were analyzed by inverted fluorescent microscopy. The clonal and proliferation ability, cel cycle and the mineralization potential were compared before and after transfection. Based on those mentioned above, we could evaluate the influence of infection on their biological characteristics.
RESULTS AND CONCLUSION:Flow cytometry results showed that rat dental pulp stem cel s expressed STRO-1 and CD146 but not CD34 or CD45. The dental pulp stem cel s could differentiate into osteoblasts and
adipocytes when cultured in specific medium for each lineage differentiation. The highest efficiency of infection and strongest fluorescence expression appeared at 48 hours of infection and MOI 50. There were no significant differences in growth ability, cel colony formation rate and cel cycle before and after transfection (P>0.05). And the alkaline phosphatase expressed positively. Infection for 48 hours at MOI 50 is optimal for transfecting dental pulp stem cel s with green fluorescent protein by a lentiviral vector, thereby providing reliable tracer method for the study of rat dental pulp stem cel s in vivo.

Objective To construct humanized monoclonal antibodies against CD 20 and check their affinity to CD 20 antigen and their anti-tumor activity.Methods Based on the computer model , human IgG1 candidates closest to rituximab in crystal structure were selected in the Protein Data Bank ( PDB) .With the selected human IgG 1 candidates as the frame , we modified and transplanted the complementarity determining region ( CDR) of rituximab .First,the target gene fragments were obtained by overlapping PCR.Then, the sequences of the light chains(L) and the heavy chains(H) were inserted in-to the pcDNA3.3 and pOptiVEC vectors.Next, the constructed clones were transfected into 293F cells through transient transfection.After a large-scale cell culture, the mAb was purified by affinity chromatography rProtein A column.The puri-ty and expression level of the humanized antibodies was tested by sodium dodecyl sulfate ( SDS)-polyacrylamide gelelectro-phoresis(PAGE).The affinity of the humanized antibodies to CD20 was assessed with Fortebio assay.Finally, the anti-tumor activity of the constructed antibodies was detected by checking the tumor growth inhibition of the nude mice transplan-ted with tumor .Results Three humanized monoclonal antibodies against CD 20 were expressed and purified successfully . In reducing SDS-PAGE, the antibodies exhibited two bands of approximately 25 ×103 and 55 ×103 , respectively.The band size of the antibodies matched the expected value.Fortebio assay revealed that the humanized antibodies could bind to CD20 with high affinity (rituximab:6.48 ×10 -9mol/L, L4H7:1.91 ×10 -9mol/L, L5H5:7.35 ×10 -10mol/L,and L5H7:1.91 ×10 -9mol/L).The tumor growth inhibition experiment showed that the anti-tumor activity of L5H7 mAb was better than that of rituximab .Conclusion Three humanized monoclonal antibodies against CD 20 have been successfully construc-ted and expressed.L5H7 mAb possesses high affinity for CD20 and a good ability to kill tumor cells.