1.Performance of transfection with a complex plasmid encoding green fluorescent protein tagged cathepsin D in researches on chronic photodamaged fibroblasts
Yue ZHENG ; Haiyan CHEN ; Qingfang XU ; Congxiu YE ; Huixian LIU ; Jinling YI ; Wei LAI
Chinese Journal of Dermatology 2015;(12):876-879
Objective To evaluate the performance of transfection with a complex plasmid encoding green fluorescent protein tagged CatD (GFP-CatD)in researches on chronic photodamaged fibroblasts. Methods Human dermal fibroblasts (HSFs)were irradiated with ultraviolet A (UVA)at 25 J/cm2 once a day for 21 consecutive days to establish a chronic photodamaged cell model. A plasmid encoding GFP-CatD was constructed and transfected into some chronic photodamaged fibroblasts (experimental group). The photodamaged HSFs receiving no treatment served as the blank control group, and those transfected with the negative plasmid encoding GFP only as the negative control group. After additional culture, fluorescence microscopy and Western-blot analysis were performed to observe and measure the expression of GFP-CatD in HSFs respectively, flow cytometry and methyl thiazolyl tetrazolium (MTT)assay to evaluate the apoptosis and proliferation of chronic photodamaged fibroblasts respectively. Results Fluorescence microscopy showed the expression of GFP-CatD in cytoplasm of chronic photodamaged fibroblasts at 96 hours after transfection with the GFP-CatD-encoding plasmid. Western-blot analysis revealed that the expression of CatD in the experimental group was 1.28 times that in the blank control group. There were no significant differences in the apoptosis rate(4.29% ± 1.30%vs. 3.03% ± 1.70% , P > 0.05)or proliferative rate (45.20% ± 4.70% vs. 43.60 ± 3.90% , P > 0.05)between the experimental group and blank control group. Conclusion CatD could be traced in chronic photodamaged fibroblasts with no changes in biological activity or cell cycle after transfection with the GFP-CatD-encoding complex plasmid.
2.Study on rat nasal absorption in situ of borneol based on single pass perfusion method.
Xiao SONG ; Shouying DU ; Yang LU ; Yong MA ; Xiaolan CHEN ; Yue WANG ; Huixian ZHANG
China Journal of Chinese Materia Medica 2011;36(18):2489-2492
OBJECTIVETo investigate the absorption characteristic of borneol.
METHODUsing single pass perfusion model, the active ingredient of borneol were detected by GC. The drug concentration, perfusion rate and pH value on the absorption of borneol were studied.
RESULTPerfusion rate on the absorption rate constants (Ka) had significant effects. Drug concentration and pH value on the absorption rate constants had no significant impact.
CONCLUSIONthe absorption of borneol is good by nasal. The absorption rate constants of borneol have no effected by drug concentration. The absorption of borneol is via a simple diffusion.
Absorption ; Administration, Intranasal ; methods ; Animals ; Bornanes ; administration & dosage ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Perfusion ; methods ; Rats
3.Study of the subcortical cerebral blood flow metabolism in patients with Parkinson′s disease using arterial spin labeling magnetic resonance imaging
Hongying ZHANG ; Jingtao WU ; Jing YE ; Wenxin CHEN ; Yao XU ; Meng WANG ; Huixian SHI ; Yue HU ; Jun XU ; Weiqiang DOU
Chinese Journal of Neurology 2019;52(9):739-744
Objective To investigate alterations of blood perfusion in subcortical regions in patients with Parkinson′s disease (PD) by three dimentional arterial spin labeling (ASL) magnetic resonance imaging (MRI). Methods Thirty patients with PD and 40 control subjects were recruited from the inpatient and outpatient of the Department of Neurology of Northern Jiangsu People′s Hospital during October 2014 to October 2016, and routine brain MRI and 3D pseudo?continuous pulse ASL were performed on all the subjects. The cerebral blood flow (CBF) maps derived from 3D ASL were coregistered to the Montreal Neurological Institute brain space. The stereo?templates of bilateral caudate nucleus, putamen nucleus, globus pallidum and thalamus from Anatomical Automatic Labeling were used as region of interest (ROI) to exstract absolute CBF values in these subcortical regions, respectively. The CBF ratio (rCBF) values represented by individual whole brain CBF divided by each of the regional CBF were also calculated in consideration of the difference between individual whole brain CBF. The CBF and rCBF values were compared respectively between groups by one?way analysis of variance. Results The subcortical CBF values (ml·100 g-1·min-1) for each ROI in PD (caudate nucleus (left: 35.32±6.47, right: 36.17±7.07), globus pallidum (left: 40.42 ± 5.83, right: 40.18 ± 5.70), putamen nucleus (left: 41.97 ± 6.12, right: 42.91 ± 6.43) and thalamus (left: 46.58 ± 7.71, right: 49.11 ± 7.10)) were significantly lower than that in the control group (caudate nucleus (left: 41.38±7.05,right: 41.63±6.85), globus pallidum (left: 45.65±8.35,right: 45.53±8.94), putamen nucleus (left: 48.49±8.78, right: 48.99±8.88) and thalamus (left: 54.32±11.94,right: 56.21±11.98), F=13.58, 10.56, 12.11, 10.06, 8.59, 8.23, 9.57, 8.30, P=0.000, 0.002, 0.005, 0.005, 0.001, 0.002, 0.003, 0.005, respectively ). The whole brain mean CBF values of each subject were also extracted and compared bewteen groups, and mean CBF values (ml·100 g-1·min-1) in PD patients (42.14±9.61) decreased significantly than those in the control group (51.59±9.67, F=16.42, P<0.01), and there was a 18.31% decrement in whole brain mean CBF in the patient group. However, rCBF values for almost all subcortical ROIs of the patients significantly increased when compared with the control group. Conclusions The decreased absolute cerebral blood perfusion involved not only subcortical regions, but also the whole brain level in the course of PD. The CBF metabolism in patients with PD may have been redistributed, with relative hyperperfusion in the subcortical brain regions contrast to the whole brain perfusion level of patients themselves.
4.Establishment and evaluation of a method for detection of ASFV antigen by doub-le-antibody sandwich ELISA
Qixuan LI ; Huixian YUE ; Yiqian JIANG ; Yanyan ZHANG ; Teng CHEN ; Shuchao WANG ; Shoufeng ZHANG ; Rongliang HU
Chinese Journal of Veterinary Science 2024;44(8):1579-1584,1592
African swine fever(ASF)is an acute and highly pathogenic hemorrhagic disease of pigs,causing huge economic losses to pig industry.In order to quantitatively detect clinical samples of ASF and inactivated ASFV antigens,the IgG of ASF positive serum was used as capture anti-body and the HRP-labeled p72 monoclonal antibody was used as detecting antibody.The standard curve was drawn with the cell-cultured ASFV,and a sandwich ELISA detection of antigen was es-tablished.The specificity,sensitivity and stability of the method were evaluated.The effects of dif-ferent inactivation methods and adjuvant addition on antigen detection were further evaluated.The results showed that the minimum detection limits of the recombinant protein and the ASFV were 0.1 mg/L and 103.7 TCID50/mL,respectively.There was no cross-reaction with five common porcine pathogenic viruses,and the coefficient variations between batches was less than 10%.The total co-incidence rate with real-time fluorescence quantitative PCR was 92%(23/25).The sensitivity of antigen detection was significantly reduced when antigen was treated by BEI inactivation,and the detection results were severely interfered by aluminum adjuvant and nano-adjuvant.In summary,the sandwich ELISA antigen detection method established is specific,sensitive,and repeatable,with a good consistency to the qPCR method,which provides an effective clinical diagnostic meth-od for ASFV antigen.
5. Study of the subcortical cerebral blood flow metabolism in patients with Parkinson′s disease using arterial spin labeling magnetic resonance imaging
Hongying ZHANG ; Jingtao WU ; Jing YE ; Wenxin CHEN ; Yao XU ; Meng WANG ; Huixian SHI ; Yue HU ; Jun XU ; Weiqiang DOU
Chinese Journal of Neurology 2019;52(9):739-744
Objective:
To investigate alterations of blood perfusion in subcortical regions in patients with Parkinson′s disease (PD) by three dimentional arterial spin labeling (ASL) magnetic resonance imaging (MRI).
Methods:
Thirty patients with PD and 40 control subjects were recruited from the inpatient and outpatient of the Department of Neurology of Northern Jiangsu People′s Hospital during October 2014 to October 2016, and routine brain MRI and 3D pseudo-continuous pulse ASL were performed on all the subjects. The cerebral blood flow (CBF) maps derived from 3D ASL were coregistered to the Montreal Neurological Institute brain space. The stereo-templates of bilateral caudate nucleus, putamen nucleus, globus pallidum and thalamus from Anatomical Automatic Labeling were used as region of interest (ROI) to exstract absolute CBF values in these subcortical regions, respectively. The CBF ratio (rCBF) values represented by individual whole brain CBF divided by each of the regional CBF were also calculated in consideration of the difference between individual whole brain CBF. The CBF and rCBF values were compared respectively between groups by one-way analysis of variance.
Results:
The subcortical CBF values (ml·100 g-1·min-1) for each ROI in PD (caudate nucleus (left: 35.32±6.47, right: 36.17±7.07), globus pallidum (left: 40.42±5.83, right: 40.18±5.70), putamen nucleus (left: 41.97±6.12, right: 42.91±6.43) and thalamus (left: 46.58±7.71, right: 49.11±7.10)) were significantly lower than that in the control group (caudate nucleus (left: 41.38±7.05, right: 41.63±6.85), globus pallidum (left: 45.65±8.35, right: 45.53±8.94), putamen nucleus (left: 48.49±8.78, right: 48.99±8.88) and thalamus (left: 54.32±11.94, right: 56.21±11.98),