Expression of Rice Gall Dwarf Virus Outer Coat Protein Gene (S8) in Insect Cells

To obtain the P8 protein of Rice gall dwarf virus (RGDV) with biological activity,its outer coat protein gene S8 was expressed in Spodoptera frugiperda (Sf9) insect cells using the baculovirus expression system.The S8 gene was subcloned into the pFastBacTM1 vector,to produce the recombinant baculovirus transfer vector pFB-S8.After transformation,pFB-S8 was introduced into the competent cells (E.coli DH10Bac) containing a shuttle vector,Bacmid,generating the recombinant bacmid rbpFB-S8.After being infected by recombinant baculovirus rvpFB-S8 at different multiplicities of infection,Sf9 cells were collected at different times and analyzed by SDS-PAGE,Western blotting and immunofluorescence microscopy.The expression level of the P8 protein was highest between 48-72 h after transfection of Sf9 cells.Immunofluorescence microscopy showed that P8 protein of RGDV formed punctate structures in the cytoplasm of Sf9 cells.

Application value of color Doppler ultrasound in the detection of hemodynamic change in patients with chronic kidney disease

Objective:To explore the application value of color Doppler ultrasound in the detection of hemodynamic change in patients with chronic kidney disease (CKD). Methods:A total of 720 patients with CKD who were admitted in our hospital from June, 2014 to May, 2016 were included in the study and divided into CKD2 group (n=242), CKD3 group (n=230), and CKD4 group (n=248) according to K/DOQI guideline. Meanwhile, 100 healthy volunteers were served as the control group. The renal parenchymal thickness, length, width, thickness, and hemodynamics by the two-dimensional ultrasound and color Doppler ultrasound in the four groups were observed. Results:The renal parenchymal thickness, length, width, and thickness in CKD2 group were reduced when compared with the control group, but the difference was not statistically significant (P>0.05);the above indicators in CKD3 group were significantly lower than those in the control group and CKD2 group (P<0.001);and the comparison of the above indicators between CKD4 group and other three groups was statistically significant (P<0.001). The comparison of hemodynamic parameters between CKD2 group and the control group was not statistically significant (P>0.05). The various hemodynamic parameters of Vs and Vd in CKD3 group were significantly lower than those in the control group and CKD2 group (P<0.001), while the various indicators of RI were significantly higher than those in the control group and CKD2 group (P<0.001). The various hemodynamic parameters of Vs and Vd in CKD4 group were significantly lower than those in the other three groups (P<0.001), while the various indicators of RI were significantly higher than those in the other three groups (P<0.001). Conclusions:Application of color Doppler ultrasound in detection of the hemodynamic change in patients with CKD can provide a reliable evidence for the disease diagnosis, condition monitoring, and prognosis evaluation, with an important application value.

Arteriolosclerosis differs from venular collagenosis in relation to cerebrovascular parenchymal damages:an autopsy-based study

Background and purpose Cerebrovascular parenchymal damage is prevalent in ageing brains;however,its vascular aetiology has not been fully elucidated.In addition to the underlying role of sclerotic arterioles,the correlation between collagenised venules has not been clarified.Here,we aimed to investigate the associations between microvascular injuries,including arteriolosclerosis and venular collagenosis,and related parenchymal damages in ageing brains,to investigate the underlying correlations.Methods We evaluated arteriolosclerosis and venular collagenosis in 7 regions from 27 autopsy cases with no history of stroke or brain tumour.The correlations between the ratio of arteriolosclerosis,venular collagenosis and the severity of cerebrovascular parenchymal damage,including lacunes,microinfarcts,myelin loss,and parenchymal and perivascular haemosiderin deposits,were assessed.Results Arteriolosclerosis and venular collagenosis became more evident with age.Arteriolosclerosis was associated with lacunes(p=0.004)and brain parenchymal haemosiderin deposits in the superior frontal cortex(p=0.024)but not with leukoaraiosis severity.Venular collagenosis was not associated with the number of lacunes or haemosiderin,while white matter generally became paler with severe venular collagenosis in the periventricular(β=−0.430,p=0.028)and deep white matter(β=−0.437,p=0.025).Conclusion Our findings imply an important role for venular lesions in relation to microvessel-related parenchymal damage which is different from that for arteriolosclerosis.Different underlying mechanisms of both cerebral arterioles and venules require further investigation.