1.Relationship between utrophin and dystrophin in muscle of patients with several nerve and muscle diseases
Wei-Min WANG ; Chuan-Qiang PU ;
Chinese Journal of Neurology 2000;0(04):-
Objective To investigate relationship between utrophin and dystrophin in muscle of patients suffered from several neurological muscular diseases.Methods Muscle biopsies of 26 cases of patients suffered from 8 categories neurological muscular diseases and 2 cases of control were analysed for utrophin and dystrophin by immunofluorescence experiments.Results In a majority of Duchenne muscular dystrophy (DMD) patients,their sarcolemma revealed absent,weak or discontinuous fluorescence for dystrophin.In non-DMD muscular dystrophies,lipid storage myopathy,myotonic dystrophy,inflammatory myopathies, neurogenic amyotrophy, polymyositis, mitochondrial encephalomyopathy, myogenic amyotrophy,immunofluorescence reactivity for dystrophin were strongly exhibited in entire sarcolemma.In normal biopsy sample,strong immunofluorescence reactivity for dystrophin was identified in entire sarcolemma,while weak and discontinuous fluorescence was identified on a minority of sarcolemma of DMD patients with severely reduced dystrophin.There was no immunofluorescence reactivity for utrophin in sarcolemma of DMD patients with moderate decreased dystrophin,non-DMD muscular dystrophies and other 6 categories of neurological muscular diseases,nor in sarcolemma of normal biopsies.Conclusions utrophin is expressed in sarcolemma of DMD patients,who have severely reduced dystrophin simultaneously. utrophin is absent in sarcolemma of other categories of neurological muscular diseases including non-DMD muscular dystrophies with normal dystrophin expression and DMD patients with moderately decreased dystrophin.
2.Experimental study of tagged fluid in CT virtual colonoscopy
Chuan LI ; Qiang FU ; Wei CHEN ; Daiquan ZHOU ; Jian WANG
Journal of Third Military Medical University 2002;0(12):-
Objective To investigate the effects of tagged fluid and substraction in CT virtual colonoscopy (CTVC) on the conspicuity of polyps and to confirm the optimal attenuation value and the optimal viewing window. Methods Polyps measured 3-10 mm were created in fresh porcine colon in vitro and submerged in saline and mixed with positive contrast medium for the CT values of 200 HU, 400 Hu, 600 HU, and 800 HU. Polyps were measured before and after substraction and compared with those in the control group. The effects of different viewing windows and different attenuation values on the measurement and conspicuity of polyps were analyzed. Results The optimal attenuation value of tagged fluid was 800 HU and the optimal viewing window was colon (-150 HU, 1 500 HU) and bone (500 HU, 2 500 HU). Conclusion The combination of tagged fluid and CT substraction can improve the conspicuity of small polyps covered by colonic fluid.
3.The protective effect and molecular mechanism of live combined Bifidobacterium and Lactobacillus tablets against acute liver failure in mice
Wei CAO ; Caiyan ZHAO ; Wei WANG ; Xin ZHAO ; Miao SU ; Yadong WANG ; Chuan SHEN
Chinese Journal of Infectious Diseases 2015;33(1):38-43
Objective To investigate the mechanism of live combined Bifidobacterium and Lactobacillus tablets in acute liver failure (ALF) treatment.Methods Ten mice were injected intraperitoneally with 3.0 g/kg D-galactosamine to establish the model of ALF and treated with live combined Bifidobacterium and Lactobacillus tablets.Protein levels of Jagged1,Notch1,Notch intracellular domain (NICD),Hes5 and the mRNA expressions of Jagged1,Notch1,Hes5 were measured via Western blot and real time-polymerase chain reaction (PCR),respectively.The protein level of CD68 was detected by immunohistochemical staining method.Meanwhile,serum alanine aminotransferase (ALT),aspartate aminotransferase (AST),interleukin (IL)-10,high mobility group protein B1 (HMGB1) and plasma lipopolysaccharide (LPS) were measured.Moreover,model group and control group were also established with 10 mice each.In vitro,RAW264.7 cells were cultured with normal mice plasma,plasma of ALF mice and plasma of treated mice,respectively.Real time-PCR and Western blot were used to determine the mRNA expressions of Jagged1,Notch1,Hes5 and proteins levels of Jagged1,Notch1,NICD,Hes5.The levels of IL-10,HMGB1 and LPS in the supernatant of RAW264.7 cells were detected as well.The total significant differences among groups were compared by one way ANOVA,and q test was used to evaluate the significance of subgroup differences.Results The levels of serum ALT,AST,HMGB1,IL10,plasma LPS,and the expressions of Jagged1,Notch1,NICD,Hes5,CD68 were higher in ALF model group than control group (all P<0.01).Compared with the ALF model group,all of these indexes could be improved in mice with live combined Bifidobacterium and Lactobacillus tablets (HMGB1:[82.6±9.7] μg/L vs [101.9±12.4] μg/L,q=6.36,P<0.01; IL-10.:[3 183±769] pg/mL vs [4 628±842] pg/mL,q=6.79,P<0.01; plasma LPS:[7.40±0.92] EU/mL vs [11.80±0.89] EU/mL,q=18.81,P<0.01; Jagged1 mRNA:5.55±0.71 vs 7.63±1.41,q=7.22,P<0.01;Jagged1 protein:0.56±0.07 vs 0.71±0.07,q=7.20,P<0.01; Notch1 mRNA:3.66±0.67 vs 7.10±0.66,q=20.06,P<0.01; Notch1 protein:0.38±0.08 vs 0.66±0.11,q=9.57,P<0.01;NICD protein:0.47±0.05 vs 0.76±0.07,q=12.68,P<0.01; Hes5 mRNA:3.94±0.68 vs 7.95± 0.71,q=22.40,P<0.01; Hes5 protein:1.04±0.12 vs 1.20±0.07,q=5.61,P<0.01; CD68 protein:5 180±610 vs 7 685 ±417,q=16.38,P<0.01).And the differences were statistically significant.After RAW264.7 cells cultured with the plasma of ALF model mice,the levels of HMGB1,IL-10 and LPS in the supernatant and the expressions of Jagged1,Notch1,NICD and Hes5 in cells were increased,whereas if RAW264.7 cells were cultured with the plasma of treated mice,indexes mentioned above were significantly decreased (all P<0.01).Conclusions Live combined Bifidobacterium and Lactobacillus tablet could prevent the occurrence and development of ALF by decreasing the plasma level of LPS,inhibiting the activation of Notch signaling pathway in macrophages and reducing the secretion of HMGB1 and IL-10.
4.The effects and molecular mechanisms of simvastatin on liver fibrosis in rats with non-alcoholic fatty liver disease
Wei WANG ; Caiyan ZHAO ; Yadong WANG ; Xian HE ; Chuan SHEN ; Wei CAO ; Dianxing SUN
Chinese Journal of Digestion 2011;31(11):739-743
ObjectiveTo investigate the effects and molecular mechanism of simvastatin in liver fibrosis model of non-alcoholic fatty liver disease (NAFLD) in vivo and hepatic stellate cell in vitro.MethodsFirstly,the rat liver fibrosis model of NAFLD was established by high-fat diet administration and intervened with simvastatin.The expression of endothelial nitric oxide synthase (eNOS),inducible nitric oxide synthase (iNOS) and Collagen Ⅰ at mRNA and protein level were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.Secondly,quiescent phenotype of LX-2 cell line was induced by promoting adipocyte differentiation medium in vitro,and then the quiescent phenotype of LX-2 cell were treated with transforming growth factor β1(TGF-β1),Nitroso-L-arginine methyl ester (L-NAME) which was NOS inhibitor,simvastatin,TGFβ1 with simvastatin,and L-NAME with simvastatin separately.The changes of eNOS,iNOS,αsmooth muscle actin (α-SMA) and Collagen Ⅰexpressions at mRNA and protein level were determined by RT-PCR and Western blot.ResultsAs modeling time extended,the expressions of eNOS in rat's liver tissue of model group at mRNA and protein level decreased gradually,however the expression of iNOS and Collagen Ⅰ at mRNA and protein level increased gradually,compared with normal control group and the differences were statistically significant (P <0.05 and 0.01).By 24weeks,the expressions of eNOS in rat's liver tissue of simvastatin group at mRNA and protein level were increased,the expression of iNOS at mRNA and protein level were decreased and the expression of Collagen Ⅰ at mRNA and protein level were decreased (all P <0.05).The expression of eNOS in rat's liver tissue of model group negatively correlated with the expression of Collagen Ⅰ at mRNA and protein level (all P <0.01).The expression of iNOS positively correlated with that of Collagen Ⅰ at mRNA and protein level (all P <0.01).In LX-2 cell culture,L-NAME inhibited the activation of LX-2,reduced eNOS and iNOS expression and increased α-SMA and CollagenⅠexpression,consistent with the role of TGF-β1.Simvastatin could directly increase the eNOS expression both in quiescent and activated LX-2 cells,decrease iNOS expression,maintain quiescent phenotype and inhibit its activation.ConclusionsSimvastatin ameliorated the genesis and progression of liver fibrosis by increasing eNOS expression in LX-2 cells and reducing iNOS,α-SMA and Collagen Ⅰ expression.
5.The study of temporal bone scanning at low-dose with 64-slice spiral CT
Heng-Tao QI ; Wei-Chang QIN ; Cheng LIU ; Dao-Cai WANG ; Chuan-Ya LIU ; Wei WANG ;
Chinese Journal of Radiology 2001;0(02):-
Objective To study the rationality and possibility of 64 slice spiral CT in the examination of the temporal bone at low dose.Methods The same CT technique and temporal bone mode as those for clinical CT were used,two cranium specimens(four ears)were scanned with Somatom Sensation 64-slice spiral CT at different mA(380,300,200,160,120,80),and muhi-planar reformation was performed.The CT dose index at different mA groups were measured by 10 cm pencil ionization chamber and head dose phantom.Four anatomic structures on axial images(subarcuate fossa,tendon of tensor tympani, facial recess,etc),four anatomic structures on coronal images(scute,crista transversa,fenestra cochleae, etc)and six anatomic structures on double oblique images(malleus,incus,stirrup bone,upper bony semicircular,etc)were chosen to evaluate and grade the reformation images among different mA groups,and to determine the minimum mA value.Ten ears of five patients were used to test the validity of the minimum mA value.Results CT radiation dose was significantly reduced from(47.8?2.7)to(20.1?2.0)raCy (P
6.Effects of astragaloside Ⅳ on electrocardiogram and action potential of ventricular myocytes in guinea pigs
Yun, ZHOU ; Jun-jie, WANG ; Wei, ZHAO ; Chuan, ZHANG ; Wei-dong, ZHANG
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(6):677-680
Objective To observe the effects of astragaloside Ⅳ on electrocardiogram (ECG) and action potential of ventricular myocytes in guinea pigs. Methods ECG was recorded in vivo and ex vivo by using conventional ECG recording method from anesthetic guinea pigs and Langendoff perfusion model of hearts. Action potentials were recorded from isolated papillary muscles of right ventricles of guinea pigs by using microelectrode techniques. Results RR interval was prolonged by Astragaloside Ⅳ in a dose-dependent manner both in vivo and ex vivo. Astragaloside Ⅳ shortened action potential duration (APD), while had no effects on resting potential, action potential amplitude and maximal rate of depolarization. Conclusion Astragaloside Ⅳ exerts a negative chronotropic effect on heart and shortens APD of cardiac myocytes, which may be involved with calcium channels.
7.Reconstruction of rabbit knee joint cartilage defect using tissues engineering method
Hongfeng JIANG ; Wei WEI ; Yun-Chuan XIE ; Rong-Rui LI ; Rao-Sheng ZHAI ; Ri-Guang WANG ;
Chinese Journal of Microsurgery 2000;0(03):-
Objective To estimate curative effect of reconstruction of rabbit knee joint cartilage defect with the homogeneitic tissue engineered cartilages.Methods The chondrocytes were isolated and collected from articular cartilages of eight New Zealand white rabbits.The tissue engineered cartilages after culturing chondrocytes and atelocollogen for two days.Cartilage defects were created in both keen joint of twenty-six rab- bits.Complexes of chodrocytes and atelocollagen was grafted into the defect of left knee joint at once as experi- mental group,and no implantation were served as control.General and histological examination were respec- tively performed in both group at four weeks and eight weeks after surgery.Results After implantation,the defects were filled with cartilaginous tissue in experiment group,while there were only tissue in control group. Histologically,defective areas were filled with chondrocytes in experiment group,but only fibroblast in control group.Conclusion The implantation of the tissue engineered cartilages contenting with chondrocytes and atelocollogen can effectively improve reconstruction of rabbit knee joint.
8.Discussion on infectious diseases teaching of resident standardized training
Yadong WANG ; Qingcheng WANG ; Yi HAN ; Yanqing CHI ; Chuan SHEN ; Wei WANG ; Caiyan ZHAO
Chinese Journal of Medical Education Research 2017;16(6):596-600
Resident standardization training is an essential way in cultivating medical professionals. It is important for subject development and talent reserve. For cultivating high-quality resident physicians with comprehensive theory, independent clinical thinking, great practical ability, and innovative scientific idea, it recourses to both optimizing training scheme and evaluation system with highly specialization and comprehensive quality. In this study, the authors proposed incorporate training patterns includingoptimiz-ing design, innovating training, and standardized assessment based on their recent experience on resident standardization training.
9.Changes of Acin1 expression in congenital cataract mouse during retinal development
De-Wei, LI ; Tao, JIANG ; Xiao-Yan, TONG ; Xiao-Chuan, WANG ; Shuang-Shuang, WANG
International Eye Science 2015;(5):767-771
?AlM: To observe the expression of Acin1 ( apoptotic chromatin condensation inducer 1 ) in congenital cataract mouse retina during development and investigate the differences of retinal apoptosis and the connection of lens and retina development between congenital cataract mouse and normal mouse.
?METHODS: There were congenital cataract mice ( 10 female and 5 male) and normal C57BL/6 mice (10 female and 5 male) . One male and two female mice were fed in the same cage randomly. The young mice were divided into two groups: congenital cataract group and normal control group. Five young mice were treated each group on 1, 5, 9, 14, 17, 21, 26, 60d. The left eyes were fixed with 4% neutral formalin to detect AClN1 protein by immunohistochemistry and retinas from right eyes were used to detect the mRNA expression of Acin1.
?RESULTS: Acin1 had sustained expression in each group. AClN1 protein gradually expressed from the ganglion cell layer, inner nuclear layer to the outer nuclear layer following retinal development. lt mainly expressed on ganglion cell layer and inner nuclear layer, but not neuroblastoma layer. AClN1 protein positive cells on P1 ~ P14d increased in normal control group, P17d reduced, after P21d positive cells of each layers decreased. The overall trend was similar in congenital cataract group with normal control group, P1 ~ P14d positive cells count was lower than normal control group, P17-P21d positive cells were flat and higher than the normal control group. Compared with the same day of the two groups, the differences except for P17, P26, P60d were significant (P<0. 05). The overall difference was statistically significant in congenital cataract group ( Fcataract=295. 07, P<0. 01);in addition to P1 and P5, P17 and P21, the differences were statistically significant ( P< 0. 05 ) compared with each other in congenital cataract group. The overall difference was statistically significant in control group (Fnormal=214. 21, P<0. 01); in addition to P1 and P5d, the difference was statistically significant ( P<0. 05) compared with each other in control group. The expression of P17d in congenital cataract group was lower compared with that of P14d in control group, the difference was statistically significant (P<0. 05). Acin1 mRNA trends of two groups were similar with AClN1 protein. Compared with the same day of the two groups, the difference was significant except for P17, P21, P60d (P<0. 05 ) . The overall difference was statistically significant in each other of the two groups ( Fcataract=522. 29, P<0. 01;Fnormal=472. 05, P<0. 01). The difference was statistically significant compared with each day in control group ( P<0. 05). Compared with all the rest of days except for P21 and P26d, the difference was statistically significant in congenital cataract group (P<0. 05).
?CONCLUSlON: Acin1 exist differential expression of time and space in mouse retina during development, congenital cataract crystal developmental disorder may affect the expression of Acin1 and retinal cell apoptosis and development.
10.Significance of natural killer cell G2D expression and activation in patients with different immune status of chronic hepatitis B virus infection
Yadong WANG ; Wei WANG ; Chuan SHEN ; Li ZHANG ; Xiaobo SHANG ; Qianfei LIANG ; Caiyan ZHAO
Chinese Journal of Infectious Diseases 2017;35(1):5-10
Objective To investigate the differences of expression and activation of natural killer (NK) cell G2D (NKG2D) in patients with different immune status of chronic hepatitis B virus (HBV) infection, and to explore the significance of NKG2D-mediated immune injury in HBV infection.Methods Fifteen chronic HBV carriers (immune tolerance),15 chronic hepatitis B (CHB, immune activation) patients, 15 HBV-related acute/subacute-on-chronic liver failure (HBV-ACLF, immune over-activation) patients were enro1led in this study from January 2010 to December 2011 in the Third Hospital of Hebei Medical University.The frequencies of NK cells and NKG2D+ NK cells in peripheral blood mononuclear cells (PBMC) were detected by flow cytometry.The NKG2D mRNA expressions were measured by real-time fluorescent quantitative polymerase chain reaction.Localization and hemi-quantitative analysis of NKG2D+ cells in liver tissue were performed by immunohistochemistry staining.Concentrations of serum interferon(IFN)-γ, tumor necrosis factor(TNF)-α, perforin and granzyme B were quantified by enzyme 1inked immunosorbent assay (ELISA).Normally distributed continuous variables were analyzed using one-way analysis of variance (ANOVA), followed by Student-Newman-Keuls q test for evaluating variances between each two groups.For non-normally distributed data or heterogeneity of variance, differences between groups were analyzed using nonparametric Kruskal-Wallis H test, followed by Nemenyi test for pairwise comparisons.Pearson chi-square test was used to analyze categorical variables.Results The percentages of NK cells in PBMC were (13.58±3.24)% in healthy controls, (5.42±2.18)% in chronic HBV carriers, (7.92±2.85)% in HBV-ACLF group and (8.43±2.92)% in CHB group.The percentage of NK cells in PBMCs was lower in each chronic HBV-infected group compared with healthy controls (F=22.04, P<0.05).The frequency of NKG2D+ NK cells in HBV-ACLF group (18.92±5.85)% was the highest, followed by CHB group (12.85±3.39)%, healthy controls (8.45±2.86)%, and chronic HBV carriers (3.36±1.05%), with the statistically significant differences between each two groups (H=46.09, P<0.01).Intrahepatic NKG2D mRNA expression and NKG2D+ cells density were highest in HBV-ACLF group (6.58±1.86 and 30.69±6.67, respectively), followed by CHB group (3.25±0.95 and 17.36±4.13, respectively) and chronic HBV carriers (0.69±0.20 and 3.16±1.24, respectively), with the statistically significant differences between each two groups (H=52.10 and 52.73 respectively, both P<0.01).The similar patterns were observed in serum IFN-γ, TNF-α, perforin and granzyme B concentrations.Conclusions NKG2D expresses variously in patients with different immune status of chronic HBV infection.Activation of NKG2D may take part in the immune pathogenesis of chronic HBV infection.