Objective To investigate changes of N-methyl-D-aspartic acid receptor 2B(NR2B) level and calcium/calmodulin-dependent protein kinase Ⅱ(CaMK Ⅱ) activities in the hippocampus of vascular dementia(VD) rats and the intervention effects of Memantine.Methods The VD rat models were established by permanent,bilateral occlusion of the common carotid arteries.The rats were randomly divided into VD group and Memantine-treated group.At 4,8,12,16 weeks after operation,the water maze test was performed to detect the ability of learning and memory of the rats.The changes of NR2B level were measured by RT-PCR.The changes of the CaMK Ⅱ activities were determined by incorporation of 32P into histone.The resoults were compared with the sham-operated group.Results Compared with the sham-operated group,the ability of learning and memory of VD group rats at each time point after operation decreased significantly(all P

The stems and branches of Hypericum petiolulatum were extracted by alcohol and liquid-liquid extraction. Seven furofuran lignans were isolated from the ethyl acetate fraction of ethanol extract of H. petiolulatum by using silica gelchromatography, Sephadex LH-20 chromatography, medium-pressure liquid chromatography and preparative HPLC. Their structures were identified by the spectroscopic methods as pinoresinol (1), medioresinol (2), 8-acetoxypinoresinol (3), epipinoresinol (4), (+)-syringaresinol (5), (+)-1-hydroxysyringaresinol (6) and erythro-buddlenolE (7). All the isolates were firstly found in H. petiolulatum. In the bioassay, compound 7 showed remarkable antioxidative activity inhibiting Fe(+2)-cystine induced rat liver microsomal lipid peroxidation with inhibitory rate 38% at a concentration of 1 x 10(-6) mol · L(-1) (positive control Vit E with the inhibitory rate of 35% at the same concentration).
Animals ; Antioxidants ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Hypericum ; chemistry ; Lipid Peroxidation ; drug effects ; Microsomes, Liver ; drug effects ; metabolism ; Molecular Structure ; Oxidative Stress ; drug effects ; Plant Stems ; chemistry ; Rats

BACKGROUND:Specific down-regulation of interleukin 1 beta (IL-1β) may al eviate the pain behaviors effectively after peripheral nervous injury. Compared with smal interference RNA (siRNA), short hairpin RNA (shRNA) could inhibit the expression of target gene more stably and efficiently. However, simple shRNA could not enter target cel s to down-regulate target gene efficiently. Adenovirus vectors have wide host range, high infection efficiency and stable expression in host cel s.
OBJECTIVE:To construct recombinant adenovirus vector expressing shRNA targeting IL-1βgene and detect its effect on the expression of target gene.
METHODS:Three siRNAs were designed on the basis of the nucleotide sequence of IL-1βobtained from NCBI and then three shRNAs (shRNA1, shRNA2 and shRNA3) were synthesized. The annealed shRNA product and adenovirus vector pHBAd/U6/GFP digested by BamH I and EcoR I were connected to construct the recombinant adenovirus vector shuttle plasmid expressing shRNA targeting IL-1β. After sequencing, HEK 293 cel s were co-transfected by the shuttle plasmid and skeleton vector, and three recombinant adenovirus vector expressing shRNA targeting IL-1β(rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3) were packaged and amplified. Rats H9C2 cel s were infected by recombinant adenovirus vector expressing shRNA targeting IL-1βand fluorescence microscope was used to observe the infection efficiency. The effect of recombinant adenovirus vector expressing shRNA targeting IL-1βon the expression of target gene was detected by western blot assay.
RESULTS AND CONCLUSION:The sequencing results showed that the sequences of three shRNAs adenovirus vector shuttle plasmid were consistent with the sequences of three designed shRNAs. rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3 were constructed successful y. rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3 could down-regulate the expression of IL-1βin rat H9C2 cel s and the down-regulation effect of rAd/shRNA2 was the most significant.

Objective To evaluate the change of left atrial function after mitral valvuloplasty in patients with mitral valve prolapse by measuring left atrial strain using two-dimensional speckle tracking echocardiography(2D-STE).Methods Study population consisted of 25 patients with mitral valve prolapse and 25 healthy subjects.High frame rate two-dimensional images were recorded from the apical four chamber view and two chamber view.Images of patients with mitral valve prolapse were respectively recorded at pre-operation,3 days and 3 months post-operation.Left atrial longitudinal strain was measured in two views using two-dimensional strain soft ware.Results ①Compared with the control group,global peak atrial longitudinal strain (PALS),peak atrial contraction strain (PACS) and atrial longitudinal strain during early diastole(ALSED) decreased (P ＜0.01) in patients with mitral valve prolapse,but global timeto-peak of peak atrial longitudinal strain(TPLS) increased(P ＜0.01).Regurgitant jet area/left atrium area (RJA/LAA) in patients with mitral valve prolapse correlated negatively with global PALS(r =-0.620,P ＜ 0.01).② Compared with preoperative group,global PALS,PACS,ALSED and TPLS decreased significantly at 3 days after surgery(P ＜0.01).Global PALS,ALSED(P ＜0.01) and PACS(P ＜0.05)increased at 3 months after surgery,but global TPLS did not differ significantly.Conclusions 2D-STE could accurately evaluate the change of left atrial function after mitral valvuloplasty.

Objective To evaluate the effects of isoflurane postconditioning on long-term cognitive function of neonatal rats with hypoxic-ischemic brain injury (HIBI).Methods Sixty 7-day-old Sprague-Dawley rats,weighing 12-16 g,were randomly divided into 4 groups (n =15 each) using a random number table:sham operation group (group Ⅰ),isoflurane postconditioning group (group Ⅱ),cerebral hypoxia-ischemia group (group Ⅲ),and isoflurane postconditioning after cerebral hypoxia-ischemia group (group Ⅳ).Brain ischemia was induced by permanent ligation of the left common carotid artery followed by inhalation of 8 ％ O2-92 ％ N2 for 2 h at 37 ℃ in Ⅲ and Ⅳ groups.In Ⅰ and Ⅱ groups,the left common carotid artery was only isolated but not ligated.The rats inhaled 1.5％ isoflurane in 30％ O2-70％ N2 for 30 min starting from 2 h of hypoxia in Ⅱ and Ⅳ groups.The rats were exposed to 30％ O2-70％ N2 for 30 min in Ⅰ and Ⅲ groups.Morris water maze test was carried out at 30-35 days after HIBI.The escape latency,swimming speed,swimming distance,the number of times the animals crossing the platform quadrant,the percentage of time spent in the platform quadrant and the percentage of swimming distance in the platform quadrant were recorded.The animals were sacrificed after Morris water maze test.The density of normal neurons in ventral posterior inferior thalamic nucleus and hippocampal CA3 region in left and right cerebral hemisphere was measured and the ratio of the density of normal neurons in the left to right cerebral hemisphere was calculated.Results Compared with group Ⅰ,the escape latency was significantly prolonged at 30-34 days after HIBI in group Ⅲ and at 31 and 34 days after HIBI in Ⅳ group,the number of times the animals crossing the platform quadrant,percentage of time spent in the platform quadrant,percentage of swimming distance in the platform quadrant,and ratio of the density of normal neurons in the left to right cerebral hemisphere were decreased at day 35 after HIBI in group Ⅲ,no significant changes were found in the number of times the animals crossing the platform quadrant,percentage of time spent in the platform quadrant,and percentage of swimming distance in the platform quadrant,and the ratio of the density of normal neurons in the left to right cerebral hemisphere was decreased at day 35 after HIBI in group Ⅳ,and no significant changes were found in the parameters mentioned above in group Ⅱ.Compared with group Ⅲ,the escape latency was significantly shortened at 31-34 days after HIBI,and the number of times the animals crossing the platform quadrant,percentage of time spent in the platform quadrant,percentage of swimming distance in the platform quadrant,and ratio of the density of normal neurons in the left to right cerebral hemisphere were increased at day 35 after HIBI in group Ⅳ.There was no significant difference in the swimming speed and swimming distance at day 35 after HIBI between groups.Conclusion Isoflurane postconditioning can improve long-term cognitive function of neonatal rats with HIBI.

Objective To evaluate the effects of isoflurane postconditioning on mitochondrial permeability transition pore (mPTP) in brain tissues of neonatal rats with hypoxic-ischemic brain injury.Methods One hundred and twenty 7-day-old Sprague-Dawley rats,weighing 12-16 g,were randomly divided into 4 groups (n =30 each) using a random number table:sham operation group (group S),isoflurane group (group I),hypoxicischemic brain injury group (group HIBI),and hypoxic-ischemic brain injury + isoflurane postconditioning group (group HI).To establish hypoxic-ischemic brain injury model in the neonatal rats,the left common carotid artery ligation was carried out,and then the rats were exposed to 8％ O2 + 92％ N2 at 37 ℃ for 2 h in HIBI and HI groups.The rats inhaled 1.5 ％ isoflurane for 30 min after the model was established in group HI.The rats only inhaled 1.5％ isoflurane for 30 min in group I.At 24 h after the model was established,10 rats taken out randomly in each group were sacrificed and brains were removed to detect mPTP opening.At 7 days after the model was established,the survival rate was recorded in the rest rats.The rats were then sacrificed and brains were removed and the right and left cerebral hemispheres were weighed separately,and the ratio between left/right cerebral hemispheres was calculated.The density of normal neurons in ventral posterior inferior thalamic nucleus and hippocampal CA3 region in the left and right cerebral hemispheres were measured and the ratios of the density of normal neurons in the left to right cerebral hemisphere were calculated.Results There was no significant difference in the survival rate between the four groups (P ＞ 0.05).Compared with group S,the ratios of the density of normal neurons in the left to right cerebral hemisphere,weight of left cerebral hemisphere,and ratio between left/right cerebral hemispheres were significantly decreased,and mPTP opening was increased in group HIBI (P ＜ 0.05),and no significant changes were found in the parameters mentioned above in group I (P ＞ 0.05).Compared with group HIBI,the ratios of the density of normal neurons in the left to right cerebral hemisphere,weight of left cerebral hemisphere,and ratio between left/right cerebral hemispheres were significantly increased,and mPTP opening was decreased in group HI (P ＜ 0.05).Conclusion The mechanism by which isoflurane postconditioning reduces hypoxic-ischemic brain injury may be related to inhibition of mPTP opening in brain tissues of neonatal rats.

Increasing attention has been focused on the antitumor activity of artemisinin derivatives in recent years, for artemisinin had been reported to have cytotoxic effects against HL-60, P388 and MCF-7 tumor cells. We report here the synthesis and evaluation for antitumor activity of a series of artemisinin-ether derivatives bearing tetrahydropyrrole, morpholine, piperidine, substituted piperidines and azoles with various linkers. Sixteen 10-O-substituted dihydroartemisinin derivatives were designed and synthesized, all of which have never been reported in literatures and whose antiproliferative effects on human breast cancer MCF-7, MCF-7/Adr and HL-60 cells were determined by MTT assay or direct cell counting. Each of these artemisinin derivatives possessed better effects than dihydroartemisinin evidently against HL-60 and MCF-7 cells growth, while less potent than doxorubicin. All target compounds exhibited significantly improved potency compared to DHA and doxorubicin on the doxorubicin-resistant MCF-7/Adr cells, so did they in their sensitive counterparts MCF-7 cells. Among them, compounds GF02, GH04 and ZH04 showed strong activity against these three cell lines growth. Further research is undergoing.

Objective To evaluate the role of interleukin?1β( IL?1β) in activation of N?methyl?D?aspartate ( NMDA) receptors in spinal dorsal horns of rats with neuropathic pain. Methods One hundred twenty?eight pathogen?free healthy adult male Sprague?Dawley rats, weighing 200-250 g, were divided into 4 groups (n=32 each) using a random number table: control group (group C), recombinant adenovirus vector group ( group rAd) , recombinant adenovirus vector shRNA group ( group rAd∕shRNA) , and recom?binant adenovirus vector shRNA plus recombinant adenovirus vector IL?1β group ( group rAd∕shRNA+rAd∕IL?1β) , receiving intrathecally injected equal volume of 0.9% sodium chloride solution, recombinant ade?novirus vector, recombinant adenovirus vector shRNA, and recombinant adenovirus vector shRNA plus re?combinant adenovirus vector IL?1β5 μl, respectively. Chronic compression of dorsal root ganglia was per?formed on 8th day after intrathecal injection. The thermal paw withdrawal latency ( TWL) was measured at 1 day before intrathecal injection ( T0 ) and at 1, 2, 3 and 5 weeks after intrathecal injection ( T1-4 ) . After measurement of the pain threshold at each time point, 8 rats selected randomly were anesthetized and sacri?ficed, and the lumbar enlargement segments of the spinal cord were harvested for determination of the ex?pression of IL?1β and phosphorylated NMDA receptor NR1 subunits at serine 896 ( pNR1S896) in spinal dorsal horns on the injured side by immunohistochemistry and Western blot. Results Compared with group C, the TWL was significantly prolonged, and the expression of IL?1βand pNR1S896 in spinal dorsal horns was significantly down?regulated at T2-4 in group rAd∕shRNA (P<0.05), and no significant change was found in the parameters mentioned above at each time point in group rAd and group rAd∕shRNA+rAd∕IL?1β( P>0.05) . Compared with group rAd, the TWL was significantly prolonged, and the expression of IL?1βand pNR1S896 in spinal dorsal horns was significantly down?regulated at T2-4 in group rAd∕shRNA ( P<0.05) , and no significant change was found in the parameters mentioned above at each time point in group rAd∕shRNA+rAd∕IL?1β (P>0.05). Compared with group rAd∕shRNA, the TWL was significantly short?ened, and the expression of IL?1β and pNR1S896 in spinal dorsal horns was significantly up?regulated at T2-4 in group rAd∕shRNA+rAd∕IL?1β ( P<0.05) . Conclusion IL?1β is involved in the activation of NM?DA receptors in spinal dorsal horns of rats with reuropathic pain.

Objective To establish CT image-based, three-dimensional finite element models of healthy tibiae and plateau-fracture tibiae, and to calculate the displacement and stress distribution of the tibial models .Methods Continuous-time tomographic images of knee joints of a healthy adult and a patient with tibial plateau fractures were obtained using multi-slice spiral CT scan , and inputed to Mimics to establish three-dimensional surface mesh models of tibiae .The models underwent global meshing procedures and material properties assignment to construct finite element models of normal and plateau-fracture tibiae in ANSYS , and the newly established models were analyzed and calculated .Results In case of applied load on the tibial plateau , the peak strains of the load-contact sites in the healthy subject and the patient presented divergent directions , with peak strains on the medial tibia in the healthy subject and lateral tibia in the patient , respectively . Equivalent stress decreased gradually down the tibiae and concentrated in the one -third of the upper and middle regions in both types of tibiae .Moreover, concentration of stress was also present in the locus of the fractured tibia .The deformation displacement gradients were more evident in the healthy tibia than in the fractured tibiae .There was significant difference in overall stress distribution between the two types of tibiae .Conclusion Three-dimensional finite element models of tibiae have been established that can demonstrate the differences in biomechanical properties between healthy and plateau -fracture tibiae, which might provide reference and guidance for orthopedic regimens .

Objective To investigate MR imaging features of femoral marrow in treated ?-thalassemia major.Methods MR imaging of the proximal femoral marrow was performed in 35 cases of ?-thalassemia major and 45 age-and sex-matched normal children as control.Coronal images of femoral marrow with the techniques of spin echo and fast field echo(FFE)were obtained.On T_1-weighted imaging the red and yellow femoral marrow were judged and marrow distribution was classified into five groups.The hemosiderosis of marrow was judged on the basis of signal intensity of marrow on FFE imaging.The marrow distribution classification and the hemosiderosis on MR imaging were correlated with clinical features.Results On FFE,marrow hemosiderosis occurred in 15 patients with a marked hypo-intensity signal and was related to the age(P=0.032).On T_1-weighted imaging,the femoral marrow in 35 patients was classified as groupⅢand IV,while the marrow distribution was groupⅠorⅡin all normal children,there was statistically significant difference(P