1.Group Ⅱ and Ⅲ metabotropic glutamate receptors agonists reverse 1-methyl-4-phenylpyridinium-induced glutamate uptake inhibition in C6 glioma cells
Fang WANG ; Honghong YAO ; Gang HU
Chinese Journal of Clinical Pharmacology and Therapeutics 2004;0(10):-
AIM: To study the effect of group Ⅱ and Ⅲ metabotropic glutamate receptors (mGluRs) agonists on 1-methyl-4-phenylpyridinium (MPP~+)-induced glutamate uptake inhibition in C6 glioma cells. METHODS: The glutamate uptake into astrocytes was measured by using radio-ligand binding assay method. RESULTS: It was shown that Group Ⅱ mGluRs agonist (2' S, 2' R, 3 ' R) -2- (2', 3 ' -dicarboxycyclopropyl) glycine (DCG-Ⅳ) (100 ?mol?L~(-1)) and Group Ⅲ mGluRs agonist L(+)-2-amino-4-phosphonobutyric acid (L-AP4) (100 ?mol?L~(-1)) significantly reversed MPP~+-induced glutamate uptake inhibition. Furthermore, the enhancement effects of DCG-Ⅳ and L-AP4 were blocked by their respective antagonists, (RS)-1 -Amino-5-phosphonoinan-1-carboxylic acid (APICA) and (RS)-?-methylserine-O-phosphate (MSOP). CONCLUSION: Group Ⅱ and Ⅲ mGluRs agonists produce neuroprotective effects by enhancing the activity of glutamate transporters.
2.Group Ⅱ and Ⅲ metabotropic glutamate receptors agonists reverse 1-methyl-4-phenylpyridinium -induced glutamate uptake inhibition
Fang WANG ; Honghong YAO ; Gang HU
Chinese Journal of Clinical Pharmacology and Therapeutics 2002;0(05):-
AIM: To study the effects of group Ⅱ and Ⅲ metabotropic glutamate receptors (mGluRs) agonists on 1-methyl-4-phenylpyridinium (MPP+) -induced glutamate uptake inhibition. METHODS: The glutamate uptake into astrocytes was measured by using radio-ligand binding assay method,and the viability of astrocytes was investigated by MTT method. RESULTS: It was shown that MPP+(150, 200 ?mol?L -1 ) inhibited glutamate uptake into astrocytes,but produced no effect on the viability of astrocytes,and the inhibition rates were 58.3 % and 70.1 %,respectively. Group Ⅱ mGluRs agonist (2'S,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl)glycine (DCG-IV) ( 0.1 ,1,10, 100 ?mol?L -1 ) and Group Ⅲ mGluRs agonist L(+)-2-amino-4-phosphonobutyric acid (L-AP4) (1,10, 100 ?mol?L -1 ) significantly reversed MPP+-induced glutamate uptake inhibition. CONCLUSION: MPP+ directly inhibits the function of glutamate transporters,and group Ⅱ and Ⅲ mGluRs agonists produce neuroprotective effects by enhancing the activity of glutamate transporters.
3.The expressions of hypoxia inducible factor -1 and angiopoietin -2 in patients with angiodysplasia related gastrointestinal bleeding and treatment with thalidomide
Honghong TAN ; Zhizheng GE ; Yunjie GAO ; Huimin CHEN ; Haiying CHEN ; Jingyuan FANG ; Wenzhong LIU ; Shudong XIAO
Chinese Journal of Digestion 2011;31(3):160-163
Objective To study the expressions of hypoxia inducible factor (HIF)-1 and angiopoietin (Ang)-2 in repeated gastrointestinal bleeding due to vascular malformation, and the efficacy of treatment with thalidomide. Methods Twenty-five patients with repeated gastrointestinal bleeding due to vascular malformation confirmed by capsule endoscopy or enteroscopy were collected and 18 subjects without severe diseases were served as controls. Ten patients with gastrointestinal vascular malformation, who received 25 mg of thalidomide 4 times daily for 4 months and were followed up for at least one year, were also enrolled. The serum samples from all participauts were detected for expressions of HIF-1 and Ang-2 using enzyme-linked immunosorbent assay (ELISA).The expressions of HIF-1 and Ang-2 were compared between angiodysplasia group and control group.The expressions of HIF-1 and Ang-2 were comparatively evaluated before and after treatment with thalidomide in treatment group. Results The expressions of HIF-1 and Ang-2 in vascular malformation group [( 113. 84 ± 26. 66 ) ng/ml and ( 652. 11 ± 140. 39) ng/ml, respectively] were significantly higher than that of control group [(43.28±17.30) ng/ml and (265.60±53.88) ng/ml,respectively, P=0. 000]. The expression of HIF-1 was positively associated with that of Ang-2. (r=0. 700, P= 0. 000). There was no difference in expressions of HIF-1 and Ang-2 before and after treatment with thalidomide (P=0. 498 and =0. 136, respectively). However, significant reduction of Ang-2 [(113. 80±73. 60) ng/ml(P=0. 003)] was found in 8 effectively treated patients after thalidomide treatment. Conclusions HIF-1 and Ang-2 might play an important role in the formation of vascular malformation. The extent of Ang-2 reduction after thalidomide treatment may be helpful in evaluating its efficacy or prognosis.
4.The potential pathogenesis of gastrointestinal vascular malformation and the potential mechanism of thalidomide in the treatment of gastrointestinal vascular malformation
Qian FENG ; Honghong TAN ; Zhizheng GE ; Yanjie GAO ; Huimin CHEN ; Jingyuan FANG ; Shudong XIAO
Chinese Journal of Internal Medicine 2012;51(5):385-389
ObjectiveTo study the pathogcncsis of gastrointestinal vascular malformation (GIVM) and the potential mechanism of thalidomide in the treatment of gastrointestinal bleeding due to GIVM.Methods We collected the surgical intestinal specimens from 10 patients who suffered from massive hemorrhage of gastrointestinal tract owning to GIVM and the normal intestinal mucosa around the lesions,as well as normal intestinal mucosa from healthy subjects.Immunohistochemical(IHC) staining was carried out to investigate the differences of angiopoietin 2 ( Ang2 ),Notch1 and delta like ligand 4 (Dll4) in the above three intestinal mucosa to find the relationship with the pathogenesis of GIVM. Human umbilical vein endothelial cells(HUVECs) were cultured with 0,25,50,100 and 200 mg/L thalidomide for 24 or 48 hours to observe their mRNA and protein expressions of Ang2,Notch1,Dll4 by real-time PCR and Western blot.ResultsBy IHC staining,more expressions of Ang2,Notch1 and Dll4 in the lesions were detected than those in the normal intestinal mucosa around the lesions and the normal intestinal mucosa in healthy people.The expressions of Ang2,Notch1 and Dll4 were significantly correlated (P =0.016,r =0.732),and the expressions of Notch1 and Dll4 were absolutely correlated ( P =0.000,r =1.000).Real-time PCR and Western blot showed that thalidomide could down-regulate the expressions of them,which were in a concentration-dependent manner.ConclusionAng2,Notch1 and Dll4 may correlate with the pathogenesis of GIVM,while thalidomide can concentration-dependently down-regulate the expression of Ang2,Notch1 and Dll4,which may be one of the mechanism that thalidomide play a therapeutic role in GIVM.
5.Porcine VHL gene cloning and construction of VHL knockdown cloned embryos.
Honghong JIN ; Jianyu WANG ; Fang WANG ; Jing MA ; Yanshuang MU ; Zhonghua LIU
Chinese Journal of Biotechnology 2013;29(6):716-725
Von Hippel-Lindau (VHL) disease is an autosomal dominant disorder and its clinical manifestation including haemangioblastomas of the central nervous system, renal cell carcinoma, haeochromocytomas, and pancreatic cyst. The deletion, mutation and promoter methylation of VHL gene can cause VHL disease. Swine is considered as an ideal model for human disease because of its physiological and anatomical similarity to human. We cloned pig VHL gene that is 2 725 bp in length. VHL highly expressed in adrenal gland, liver, pancreas, heart and testis. We designed 5 shRNAs and screened the most effective interference RNA fragment with a knockdown efficiency of 72%. Porcine embryonic fibroblasts stably transfected with pGenesil-shRNA vector were used as donor cells for nuclear transfer and there was no significant difference of embryo development compared with the control group. Moreover, VHL was efficiently knocked-down with efficiency of 71% in porcine cloned blastocyst, these results lay a solid foundation for constructing the VHL knock-down model of pig.
Animals
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Cloning, Molecular
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Disease Models, Animal
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Embryo, Mammalian
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Gene Knockdown Techniques
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Swine
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Von Hippel-Lindau Tumor Suppressor Protein
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genetics
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von Hippel-Lindau Disease
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genetics
6.Desmoplakin expression silencing affects cardiac voltage-gated sodium channel Nav1.5 in HL-1 cells.
Qianhuan ZHANG ; Chunyu DENG ; Fang RAO ; Xiaoying LIU ; Liping MAI ; Jiening ZHU ; Honghong TAN ; Shulin WU
Journal of Southern Medical University 2013;33(7):983-989
OBJECTIVETo investigate the association of desmoplakin with the distribution and function of Nav1.5 by RNA silencing technology in HL-1 cells.
METHODSHL-1 cells with desmoplakin expression suppression by RNA silencing were examined for desmoplakin and Nav1.5 protein expressions by Western blotting, and the distribution and co-location of desmoplakin and Nav1.5 protein were detected by immunofluorescence staining. Patch-clamp recording was applied to analyze the changes in whole-cell sodium current after desmoplakin silencing.
RESULTSCompared with the untreated group and negative control group, the cells with desmoplakin silencing showed obviously reduced expressions of desmoplakin and Nav1.5 proteins. Co-localization of desmoplakin and Nav1.5 was detected at cell-cell contact in untreated and control conditions, and desmoplakin expression silencing induced a drastic redistribution of Nav1.5 with decreased peak current density (156.3∓6.2 vs 41.8∓3.1, n=6, P<0.05), a shift in voltage dependence of steady-state inactivation (-42 mV vs -61 mV, n=5, P<0.05), and prolonged time of recovery from inactivation.
CONCLUSIONDesmoplakin silencing caused redistribution of Nav1.5 protein and also changes in its electrophysiological properties in HL-1 cells.
Animals ; Cell Line ; Desmoplakins ; genetics ; metabolism ; Gene Silencing ; Mice ; Mutation ; Myocytes, Cardiac ; metabolism ; NAV1.5 Voltage-Gated Sodium Channel ; metabolism
7.Identification of latent class of sleep quality among breast cancer patients during chemotherapy and differences on quality of life based on latent profile analysis
Zhaohui GENG ; Qiong FANG ; Nan ZHANG ; Danwei SHEN ; Lizhi ZHOU ; Xianjing MENG ; Honghong TAN ; Changrong YUAN
Chinese Journal of Practical Nursing 2022;38(6):431-437
Objective:To identify the classification characteristics and quality of life (QOL) of breast cancer (BC) patients during chemotherapy, so as to provide basis for improving the sleep and QOL of this group.Methods:A cross-sectional investigation was completed among 421 BC patients in 5 tertiary hospitals in Shanghai, Wuhan, Tangshan and Nanning in 1-12 months of 2016 using validated instruments including self-made general information questionnaire, Pittsburgh Sleep Quality Index (PSQI) and Functional Assessment of Cancer Therapy-Breast (FACT-B).Results:Four latent class of patients were identified through latent profile analysis (LPA), named by badly worse sleep quality(SQ) (C1, n=23), medium-SQ with difficulty to fall asleep (C2, n=127), medium-SQ with worse sleeping process (C3, n=30), none sleep disorders (C4, n=241). Total points of SQ among C1-C4 had significant difference ( χ2 value was 309.28, P<0.05). Age, BMI, job status, whether had surgery and course of chemotherapy between classes had statistically significant differences ( χ2 values were 9.57-25.28, all P<0.05). It had significant difference between C2 and C3, C2 and C4, C3 and C1, C3 and C4 on QOL ( χ2 values were 5.96-52.73, all P<0.05). Conclusion:SQ of BC patients during chemotherapy has heterogeneity among population. Different features of SQ of BC patients have different performance on QOL. Health professionals should keep an eye on patients with features of older age, high BMI, in job status, already received surgery and during early-stage chemotherapy, provide personal nursing intervention to improve SQ and QOL.
8.Association between N-glycans of immunoglobulin G and central obesity
Di LIU ; Jing DONG ; Zhongyao ZHAO ; Xi CHU ; Xinwei YU ; Hao WANG ; Qing GAO ; Honghong FANG ; Yang SUN
Chinese Journal of Health Management 2017;11(6):525-530
Objective To explore the association between immunoglobulin G (IgG) glycosylation and central obesity in a Chinese population. Methods We recruited 517 Chinese individuals from a community-based cohort in the Xicheng District, Beijing, China. The height, weight, waist circumference, and hip circumference were measured, and blood samples were collected. Plasma IgG N-glycome was determined using hydrophilic interaction liquid chromatography (HILIC). Central obesity was determined using the waist-to-hip ratio (WHR). Men and women with a WHR>0.90 and >0.80, respectively were assigned to the central obese group,and those with a WHR≤0.90(men)and≤0.80(women)were assigned to the normal group.The correlation between WHR and glycosylation was determined using the standardized regression coefficient,and P<0.05 was considered statistically significant.Results The study included 517 participants,with an average age of(47.81±5.58)years(range:27-68 years),and 164(31.70%)of them were men. The central obese and normal groups included 205 (39.65%) and 312 (60.35%) participants, respectively. The average age for the central obese group (49.21 ± 5.70) years was higher than that for the normal group (46.89 ± 5.30) years, and the difference was statistically significant (t=-4.73, P=0.001). The two groups showed no significant differences in the sex of the participants(χ2=0.34,P=0.558).Twenty-four N-glycan traits(GP1-GP24)were directly measured,and because of the high variability among the repeated measurements and the unknown structures, GP3 and GP20 were excluded from the analysis. The peak contents for GP4,GP8,GP14,and GP18 were higher than the others and accounted for about 50% of all the peak contents. IgG GP10 (b'=0.076, P=0.024) positively correlated with WHR, whereas IgG GP12, GP17, GP22 negatively correlated with WHR(b'=-0.076,-0.081,-0.080,all P<0.05).Additionally,the IgG GP12, GP17,and GP22(P=0.001,0.004 and 0.033,respectively)were significantly lower in the participants of the central obese group,compared to those of the normal group. Conclusion The loss of galactose and sialic acid,as well as the addition of fucose to IgG N-glycan,correlate with increased WHR.
9.Platelet antibody of blood donors in Suzhou
Li DONG ; Ming FANG ; Yujue WANG ; Honghong HE ; Hongmei WANG ; Yezhou CHEN ; Feiran WU ; Shengbao DUAN ; Longhai TANG
Chinese Journal of Blood Transfusion 2022;35(8):795-799
【Objective】 To study the incidence and specificity of platelet antibody in blood donors in Suzhou, analyze the distribution characteristics of platelet antibody in blood donors in this area, and explore the significance of platelet antibody detection in blood donors to reduce the adverse reactions toplatelet transfusion in clinical. 【Methods】 Platelet antibody detection was performed in 2178 blood donors in this area by solid-phase immunosorbent assay. The antibody specificity of the positive samples was analyzed by commercial kit, and the anti-CD36 antibody positive samples were further identified by flow cytometry and gene sequencing. 【Results】 Twelve positive samples were detected by platelet antibody screening, with a positive rate of 0.55%(12/2 178), including 5 males (0.33%, 5/2 178)and 7 females(1.06%, 7/2 178). Among the positive samples, anti-HLA-Ⅰ antibody was identified in 2 cases, anti-CD36 antibody in 1 case, and the antibody specificity was not identified in the other 9 cases. In one case, the positive rate of anti-HLA-Ⅰ antibody PRA was 31.31%(31/ 99), which was mainly specific to anti-B15, anti-B35 and anti-B40. The positive rate of anti-HLA-Ⅰ antibody PRA in the other case was 45.45%(45/ 99), which was mainly specific to anti-A2, anti-A11, anti-A24, anti-A29, anti-A33, anti-A66, anti-B15 and anti-B35. The blood donor with anti-CD36 antibody was type I CD36 deficiency, and 329_330delAC mutation occurred in exon 5. 【Conclusion】 Through antibody screening and specificity identification, the positive rate of platelet antibody in females was significantly higher than that in males(P<0.05). In addition to the common anti-HLA-I antibodies, anti-CD36 antibody was also detected in type I CD36 deficient blood donor. Therefore, the detection of platelet antibodies in blood donors is of certain clinical significance to reduce the adverse reactions to blood transfusion caused by antibodies in platelet products.