1.Sequence analysis of beta 1,3-galactosyltranferase specific molecular chaperone Cosmc gene coding region in IgA nephropathy patients
Chinese Journal of Tissue Engineering Research 2007;0(21):-
BACKGROUND: Pathogenesy of immunoglobulin A nephropathy (IgAN) is not clear up to now. Present research has verified that the key pathogenetic pathway is abnormalities of IgA1 molecular O-glycosylation induced by decrease of ?1,3-galactosyltranferase activity in IgA1 hinge region of IgAN patients. Prophase study by the authors supposed that the key of IgAN O-glycosylation abnormality might be due to the decrease of ?1,3-galactosyltranferase specific molecular protein chaperone Cosmc in B lymphocyte of peripheral blood in IgAN patients. OBJECTIVE: To measure DNA sequence of ?1,3-galactosyltranferase specific molecular chaperone in coding region of Cosmc gene in IgAN patients,and compared with the sequence of Gene Bank. DESIGN: Case-controlled observation. SETTING: Department of Nephrology,West China Hospital,Sichuan University. PARTICIPANTS: Totally 27 IgAN patients and 10 non-IgAN patients were recruited in Department of Nephrology of West China Hospital of Sichuan University from November 2005 to August 2006,and five normal controls were included in this study. All the subjects knew the fact and agreed to participate in the experiment. METHODS: The experiment was performed at the State Key Laboratory of Biotherapy of Sichuan University. 2 mL peripheral venous blood of all the samples were taken into heparin sodium anticoagulated tubes,from which total genomic DNA were extracted by phenol/chloroform precipitation method. Concentration of DNA was determined by ultraviolet spectrophotometer. The polymerase chain reaction (PCR) was used to amplify the coding region of ?1,3-galactosyltranferase specific molecular chaperone Cosmc gene in all the subjects and direct sequencing was done in PCR products of each subjects. The results of all the sequencing were compared with Gene Bank one by one. MAIN OUTCOME MEASURES: Amplification findings and sequencing of coding region of ?1,3-galactosyltranferase specific molecular chaperone Cosmc gene by PCR. RESULTS: ①Coding region of Cosmc gene located at 257-1 213,and amplified Cosmc gene was 1 247 bp. ②The sequence of Cosmc gene coding region was similar in IgAN patients,non-IgAN patients and normal controls,and no difference of gene sequence was noticed in all the result sequences as compared with the Gene Bank registered sequence. CONCLUSION: No abnormal sequence is found in coding region of Cosmc gene in IgAN patients,suggesting that this coding region probably is not associated with the abnormalities of IgA1 O-glycosylation in IgAN.
2.Sequence analysis of beta 1, 3-galactosyltranferase specific molecular chaperone Cosmc gene coding region in IgA nephropathy patients
Xiang ZHONG ; Wei QIN ; Junming FAN
Chinese Journal of Tissue Engineering Research 2007;11(21):4251-4254
BACKGROUND: Pathogenesy of immunoglobulin A nephropathy (IgAN) is not clear up to now. Present research has verified that the key pathogenetic pathway is abnormalities of IgA1 molecular O-glycosylation induced by decrease of β1, 3-galactosyltranferase activity in IgA1 hinge region of IgAN patients. Prophase study by the authors supposed that the key of IgAN O-glycosylation abnormality might be due to the decrease of β1, 3-galactosyltranferase specific molecular protein chaperone Cosmc in B lymphocyte of peripheral blood in IgAN patients.OBJECTIVE: To measure DNA sequence of β1, 3-galactosyltranferase specific molecular chaperone in coding region of Cosmc gene in IgAN patients, and compared with the sequence of Gene Bank.DESIGN: Case-controlled observation.SETTING: Department of Nephrology, West China Hospital, Sichuan University.PARTICIPANTS: Totally 27 IgAN patients and 10 non-IgAN patients were recruited in Department of Nephrology of West China Hospital of Sichuan University from November 2005 to August 2006, and five normal controls were included in this study. All the subjects knew the fact and agreed to participate in the experiment.METHODS: The experiment was performed at the State Key Laboratory of Biotherapy of Sichuan University. 2 mL peripheral venous blood of all the samples were taken into heparin sodium anticoagulated tubes, from which total genomic DNA were extracted by phenol/chloroform precipitation method. Concentration of DNA was determined by ultraviolet spectrophotometer. The polymerase chain reaction (PCR) was used to amplify the coding region of β1, 3-galactosyltranferase specific molecular chaperone Cosmc gene in all the subjects and direct sequencing was done in PCR products of each subjects. The results of all the sequencing were compared with Gene Bank one by one.MAIN OUTCOME MEASURES: Amplification findings and sequencing of coding region of β1, 3-galactosyltranferase specific molecular chaperone Cosmc gene by PCR.RESULTS: ①Coding region of Cosmc gene located at 257-1 213, and amplified Cosmc gene was 1 247 bp. ②The sequence of Cosmc gene coding region was similar in IgAN patients, non-IgAN patients and normal controls, and no difference of gene sequence was noticed in all the result sequences as compared with the Gene Bank registered sequence.CONCLUSION: No abnormal sequence is found in coding region of Cosmc gene in IgAN patients, suggesting that this coding region probably is not associated with the abnormalities of IgA1 O-glycosylation in IgAN.
3.Association between peripheral monocyte CD36 expression and rheumatoid arthritis
Youfan PENG ; Yang XIANG ; Yesheng WEI ; Shimao ZHONG
Chinese Journal of Rheumatology 2016;20(7):441-445
Objective To investigate the association between peripheral monocyte CD36 expression and rheum-atoid arthritis (RA).Methods A total of 108 patients with RA and 108 healthy individuals were included between 2014 and 2015.Disease Activity Score (DAS)28-erythrocyte sedimentation rate (ESR) and DAS28-C reactive protein (CRP) were used to estimate disease activity in patients with RA,and monocyte CD36 fluorescence intensity was determined by using flow cytometry.Student's t test or Mann-Whitney U test was used to compare the differences between the two groups.The categorical data was compared using x2 test.Correlations between variables were analyzed with Spearman correlation,Logistic regression analysis was used to identify risk factors associated with RA,we used receiver operating characteristic (ROC) curve to estimate the performance of relevant variables for RA.Results Low density lipoprotein cholesterin (LDL-C) [(2.8±0.8) mmol/L,(2.6±0.6) mmol/L,t=-2.046,P=0.042],ESR[25.5(18.00,40.00) mm/1 h,10.00(4.60,10.00) mm/1 h,Z=-12.786,P<0.01],CRP [15.9 (6.71,25.43) mg/L,2.28 (1.63,4.60) mg/L,Z=-7.980,P<0.01] and monocyte CD36 expression [(0.44±0.22)×109/L,(081 ±0.26)×109/L,t=1 1.159,P<0.01] was statistically different between the two groups.Of note,monocyte CD36 fluorescence intensity was negatively correlated with disease activity in patients with moderate and severe disease (r=-0.146,P=0.036;r=-0.259,P=0.007).In stepwise Logistic regression analysis,monocyte CD36 fluorescence intensity was independently associated with RA [OR=0.803,P=0.002,95%CI (0.791,0.820)].ROC curve analysis showed a significant performance with area under curve of 0.855 [95%CI (0.807,0.904);P<0.01],the sensitivity and specificity were 80.6% and 73.1%,respectively,the cutoff values was 0.570.Conclusion Monocyte CD36 fluorescence intensity was independently associated with RA in stepwise Logistic regression analysis.A correlation between monocyte CD36 fluorescence intensity and DAS28-CRP,DAS28-ESR was observed in RA patients.
4.In vitro effects of acitretin on the apoptosis and expressions of insulin-like growth factor binding protein 7 and vascular endothelial growth factor in HaCaT cells
Fengmei XIANG ; Zhiping WEI ; Liansheng ZHONG ; Qing YANG ; Yanqun LIU
Chinese Journal of Dermatology 2014;47(7):494-498
Objective To investigate the in vitro effects of acitretin on the apoptosis and expressions of insulin-like growth factor binding protein 7 (IGFBP7) and vascular endothelial growth factor (VEGF) in HaCaT cells.Methods Cultured HaCaT cells were treated with various concentrations (10-5,10-64,10-7,10-8 mol/L) of acitretin for various durations,with those cultured in acitretin-free medium serving as the control group.Then,CCK-8 assay was performed to evaluate the proliferation of cells after 24-,48-and 72-hour treatment,flow cytometry to detect the apoptosis of HaCaT cells,and Western blot and reverse transcription-PCR to quantify the protein and mRNA expressions of IGFBP7 and VEGF in HaCaT cells,respectively,after 48-hour treatment.Statistical analysis was carried out by one-way analysis of variance and Pearson correlation analysis.Results The proliferation of HaCaT cells was inhibited by the treatment with acitretin,and the inhibitory effect increased with the elevation of concentration and prolongation of treatment duration of acitretin.A significant decrease was observed in the proliferative activity of HaCaT cells treated with acitretin of 10-8 mol/L for 48 hours,and when the concentration of acitretin was 10-5 mol/L,the proliferation of HaCaT cells was inhibited by 39.94% ± 2.27% and 49.77% ± 1.87% at 48 and 72 hours respectively,compared with the control cells.The HaCaT cells treated with acitretin of 10-5 mol/L for 48 hours showed a significant elevation in apoptosis rate (7.617% ± 0.767% vs.1.803% ± 0.313%,P < 0.05),IGFBP7 protein and mRNA expressions (0.939 ± 0.040 vs.0.436 ± 0.013,0.872 ± 0.079 vs.0.190 ± 0.056,both P < 0.05),but a significant reduction in VEGF protein and mRNA expressions (0.213 ± 0.032 vs.0.798 ± 0.036,0.274 ± 0.041 vs.0.933 ± 0.054,both P < 0.05) in comparison to the control cells.Conclusions Acitretin can induce the apoptosis of HaCaT cells,and up-regulate IGFBP7 but down-regulate VEGF expressions in HaCaT cells at protein and mRNA levels.
5.Anti-inflammation and anti-oxidation effects of recombinant human superoxide dismutase on acute lung injury induced by meconium aspiration in infant rats.
Mei-ping LU ; Li-zhong DU ; Wei-zhong GU ; Zheng-zhu YU ; Xiang-xiang CHEN ; Zhong-sheng YU
Journal of Zhejiang University. Medical sciences 2005;34(1):55-59
OBJECTIVETo investigate the anti-inflammation and anti-oxidation effects of recombinant human CuZn superoxide dismutase(rhSOD) on acute lung injury (ALI) induced by meconium aspiration in rats.
METHODS1 ml/kg of 20% human newborn meconium suspension was intratracheally (IT) administrated to induce the model of ALI in 32 male Sprage-Dawley rats, and the animals were then randomized to 4 groups: 3 treatment groups with IT administration of 5, 10 and 20 mg/kg rhSOD dissolved in 1 ml/kg saline and the control group with IT administration of 1 ml/kg saline. The animals were killed after 24 h of treatments. The measurements included lung tissue wet/dry ratio, broncho-alveolar lavage fluid (BALF) protein, BALF protein/plasma protein (pulmonary permeability index, PPI),lung myeloperoxidase (MPO) and superoxide dismutase (SOD) activity, nitric oxide (NO) and 8-isoprostane levels. Lung injury score was also evaluated.
RESULTSCompared with the control group, pulmonary MPO activity, NO and 8-isoprostane levels were significantly decreased and SOD activity was markedly increased in all rhSOD treatment groups (P<0.05 or 0.01). Compared with the rhSOD 5 mg/kg group, pulmonary 8-isoprostane level was further low in the rhSOD 20 mg/kg group(P=0.01). Lung injury score was decreased in rhSOD 20 mg/kg group (P<0.05). But there were no statistically differences in lung wet/dry, BALF protein and PPI among all groups.
CONCLUSIONThe results suggest that a single IT dose of 5,10 or 20 mg/kg rhSOD can prevent lung damages in rats with ALI following meconium aspiration.
Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Antioxidants ; pharmacology ; Humans ; Lung ; drug effects ; pathology ; Male ; Meconium ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; pharmacology ; Superoxide Dismutase ; pharmacology
6.Recent advances in treatment of glial scar-induced inhibition of nerve regeneration.
Jian-Xiang ZHANG ; Wei-Wei HU ; Zhong LIU
Journal of Zhejiang University. Medical sciences 2009;38(6):639-643
Glial scar formed by central nervous system (CNS) injury is the main inhibitory barrier of nerve regeneration. How to promote axonal regeneration after injury,how to accelerate neural network reconstruction and how to improve brain function recovery have become a hot problem to be solved in the field of neuroscience. This article focuses on the recent advances of therapeutic strategies for axonal regeneration.
Animals
;
Astrocytes
;
pathology
;
Brain Injuries
;
pathology
;
physiopathology
;
Cicatrix
;
prevention & control
;
Humans
;
Nerve Regeneration
;
Neuroglia
;
pathology
;
Neuronal Plasticity
;
physiology
;
Neurons
;
physiology
;
Proteoglycans
;
metabolism
;
Spinal Cord Injuries
;
pathology
;
physiopathology
7.Influence of different processing methods on Lonicerae Japonicae Flos from Donghai cultivation base in Jiangsu province.
Yong-Xiang WANG ; Yong LUO ; Juan SHEN ; Yi-Fei ZHOU ; Zheng-Zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2014;39(14):2665-2669
To study the impact of five different origin processing methods, namely natural drying, drying in baking shop, drying by microwave heating, drying in drum and drying with sulphur fumigation, on the quality of Lonicerae Japonicae Flos from Donghai cultivation base in Jiangsu Province, with the contents of chlorogenic acid and galuteolin and the similarity in HPLC fingerprints as the evaluation indicators. The results showed that different origin processing methods had significant impact on the content of chlorogenic acid and the similarity in HPLC fingerprints, but with no significant difference on the content of galuteolin. By means of drying by microwave heating and drying in drum, the samples showed higher contents of chlorogenic acid, respectively 3.67% and 3.39%. The similarities of HPLC fingerprints were 0.815 and 0.793, respectively. By means of the drying in baking shop and the drying with sulphur fumigation, the contents of chlorogenic acid in the samples were 2. 87% and 2. 53% , respectively. The similarities of HPLC fingerprints were 0.964 and 0.765, respectively. The lowest content of chlorogenic acid in naturally dried samples was 1.92%. The similarity of HPLC fingerprints was 0.940. According to the findings as well as the internal control standards for Lonicerae Japonicae Flos herbs of Jiangsu Kanion Pharmaceutical Co. , Ltd. , the optimum processing method for Lonicerae Japonicae Flos from Donghai cultivation base was the drying in baking shop. This study provided a theoretical basis for determining the processing method for Lonicerae Japonicae Flos from Donghai cultivation base of Jiangsu Province.
China
;
Desiccation
;
methods
;
Drugs, Chinese Herbal
;
chemistry
;
Lonicera
;
chemistry
;
growth & development
;
Quality Control
8.Intravenous drug abuse-related infective endocarditis: report of an autopsy case.
Wei-xiang ZHONG ; Dong-ping TIAN ; De-qing WU ; Min SU
Chinese Journal of Pathology 2010;39(6):421-422
Adult
;
Aortic Valve
;
microbiology
;
pathology
;
Autopsy
;
Brain
;
microbiology
;
pathology
;
Endocarditis, Bacterial
;
complications
;
microbiology
;
pathology
;
Female
;
Heart Ventricles
;
microbiology
;
pathology
;
Humans
;
Mitral Valve
;
pathology
;
Sepsis
;
complications
;
microbiology
;
pathology
;
Substance Abuse, Intravenous
;
complications
;
microbiology
;
pathology
;
Young Adult
9.Causes of reduced expression of Cosmc gene in IgA nephropathy patients
Wei QIN ; Xiang ZHONG ; Yingjuan ZHANG ; Chunyu TAN ; Liehuan YANG ; Junming FAN
Chinese Journal of Nephrology 2008;24(6):377-381
Objective To clarify whether the Cosmc gene down-regnhtion in lgA nephropathy (IgAN) patients is resulted from genetic disorders or external suppressions. Methods Forty IgAN patients, 16 non-IgAN glomerulonephritis patients and 21 healthy controls were enrolled in the study. Genomie DNA was extracted and then Cosmc gene was amplified and sequenced. Peripheral B lymphoeytes were isolated and cultured with RPMI-1640 alone or plus lipopolysaecharide (LPS). The Cosmc mRNA expression levels at baseline, after RPMI-1640 culture or RPMI-1640+LPS treatment were measured respectively by real-time RT-PCR. Results (1) Only 2 missense mutations and 2 silent mutations were detected in coding frame region of Cosine gene in 2 IgAN patients. (2) The baseline Cosmc gene expression level was significantly lower in IgAN patients (31% of that in healthy controls) than that in healthy controls. (3) Relative quantification PCR indicated that Cosmc mRNA expression level was significantly increased (219% of baseline) after RPMI-1640 culture, and treatment of LPS could strongly inhibit this effect. (4) The Cosmc gene expression of healthy control was not affected by RPMI-1640 or LPS. Conclusion It is not genetic disorders but external suppression to cause the down-regulation of Cosmc gene mRNA expression in IgAN.
10.Comparison of clinicopathologic features of breast carcinoma in Shanghai Han and Xinjiang Uygur women
Fangfang ZHONG ; Wei ZHANG ; Chenghui WANG ; Xinxia LI ; Ruohong SHUI ; Wentao YANG ; Xiang DU
China Oncology 2014;(1):21-28
Background and purpose: Breast cancer is a group of heterogeneous diseases which has racial disparities. Our study was to elucidate the clinicopathologic features of breast carcinoma in Shanghai Han and Xinjiang Uygur women and to analyze the racial differences. Methods: In this study, 125 cases of breast invasive ductal carcinoma of Shanghai Han women and 85 cases of Xinjiang Uygur women were collected. The clinical stage was analyzed. Histological grading was observed. Immunohistochemical staining of ER, PR, HER-2, CK5/6, CK14, EGFR, Ki-67 was performed. Molecular subtypes were studied. Results:The average age of onset of breast cancer in Xinjiang Uygur women was younger than in Shanghai Han women (P<0.05), and Xinjiang Uygur women were more likely to be diagnosed at less than 35 years old (P<0.01). The proportion of stageⅠwas higher in Shanghai Han women (20.0%vs 8.2%), while the proportion of stageⅢwas higher in Xinjiang Uygur women (50.6%vs 27.2%) (P<0.01). The proportion of grade 2 was higher in Shanghai Han women (67.2% vs 43.5%), while the proportion of grade 3 was higher in Xinjiang Uygur women (47.1%vs 31.2%) (P<0.01). The proportion of luminal A subtype was higher in Shanghai Han women (36.8%vs 18.3%), while the proportion of basal-like subtype was higher in Xinjiang Uygur women (29.6%vs 12.0%) (P<0.01). The molecular subtype was associated with race and histological grade (P<0.05).Conclusion:There are racial differences in clinicopathologic features of breast carcinoma between Shanghai Han and Xinjiang Uygur women.