2.Increased susceptibility of recombinant type 2A von Willebrand factor mutant A1500E to proteolysis by ADAMTS13.
Jing-yu ZHANG ; Jian SU ; Zhen-ni MA ; Ning-zheng DONG ; Ying-chun WANG ; Chang-geng RUAN
Chinese Journal of Hematology 2012;33(3):169-172
OBJECTIVETo investigate the susceptibility of von Willebrand factor (VWF) type 2A mutant A1500E to proteolysis by metalloprotease ADAMTS13 and to provide the direct supports for the pathogenesis of VWF mutation A1500E responsible for von Willebrand disease (VWD) type 2A.
METHODSRecombinant wild-type VWF (WT-VWF) and A1500E mutant VWF transiently expressed on transfected HeLa cell lines. Expression media were collected and concentrated, then cleaved directly by recombinant ADAMTS13 (rADAMTS13). Compared with WT-VWF, the susceptibility of A1500E mutant VWF to proteolysis by ADAMTS13 was analyzed using SDS-agarose gel VWF multimers analysis.
RESULTSIn vitro the expression of VWF:Ag in the supernatants of WT-VWF and A1500E mutant VWF were 1.10 U/ml and 0.78 U/ml, respectively, while VWF:Ag in cells lysates of A1500E mutant VWF was 90.6% of that of WT-VWF. The SDS-agarose gel VWF multimers analysis showed that there were no differences between WT-VWF and A1500E mutant VWF. The A1500E mutant VWF could be efficiently cleaved by ADAMTS13 under static condition without denaturants such as urea and guanidine HCl. VWF multimeric analysis showed that high and intermediate molecular weight multimers dramatically decreased while low molecular weight multimers obviously increased. Conversely, WT-VWF could not be cleaved by ADAMTS13 under the same condition.
CONCLUSIONThe A1500E mutation resulted in VWF more susceptible to ADAMTS13-dependent proteolysis, which belonged to VWD type 2A group 2 mutation.
ADAM Proteins ; genetics ; metabolism ; ADAMTS13 Protein ; Genotype ; HeLa Cells ; Humans ; Hydrolysis ; Mutation ; Recombinant Proteins ; genetics ; metabolism ; von Willebrand Disease, Type 2 ; genetics ; metabolism ; von Willebrand Factor ; genetics
3.Effects of Huangqi on phagocytic activity of peritoneal macrophage of mice.
Kang-jian NING ; Xiang-chun RUAN ; Jin-fang LU ; Yu-xin CHENG
China Journal of Chinese Materia Medica 2005;30(21):1670-1672
OBJECTIVETo study the effects of Huangqi decoction (HQD) on phagocytic activity of peritoneal macrophage of mice.
METHODOne hundred Kunming mice, whose weight varied from 18 g to 22 g, were selected and divided into 10 groups randomly in eluding contrast group, groups conducted at different doses of HQD by ig, groups conducted in various ways of taking medicine, and groups conducted with comparative treat combining Huangqi and Dexamethasone. Mice in every group were taken medicine one time daily for 6 days.
RESULTAmong the groups treated at different doses of HQD, phagocytic rate and phagocytic index of mice, which were taken HQD by ig at high, middle, and low doses, were significantly higher (P < 0.01) than that of mice in contrast group, at the same time the effect in group with high dose was the best. Among the groups treated in various ways of taking medicine, phagocytic rate of the ip group was significantly better (P < 0.01) than that of the sc group and that of the ig group respectively, but there was not significant difference (P > 0.05) of phagocytic index among them. Among the groups combining Huangqi and Dexamethasone, Huangqi could antagonize the immunosuppressive effect of Dexamethasone obviously (P < 0.01).
CONCLUSIONHQD at different doses and with various ways of taking medicine could improve phagocytic activity of peritoneal macrophage of mice at different degree, and could antagonize the immunosuppressive effect of Dexamethasone.
Animals ; Astragalus membranaceus ; chemistry ; Dexamethasone ; antagonists & inhibitors ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Female ; Macrophages, Peritoneal ; drug effects ; physiology ; Male ; Mice ; Phagocytosis ; drug effects ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation
4.Changes of Kupffer cells during tree shrew chronically infected with hep-atitis B virus
Ping RUAN ; Jian XIAO ; Chun YANG ; Jianjia SU ; Chao OU ; Ji CAO ; Chengpiao LUO ; Yanping TANG ; Hong QIN ; Wen SUN ; Yuan LI
Chinese Journal of Pathophysiology 2014;(6):1076-1081
AIM:To explore the changes and significance of Kupffer cells in the process of tree shrew chroni -cally infected with hepatitis B virus (HBV).METHODS:The animals were divided into 3 groups.Group A consists of 6 tree shrews that were identified as persistently infected with HBV;group B consists of 3 tree shrews that were suspected as persistently infected with HBV;group C consists of 4 tree shrews that were not inoculated with HBV and were applied as normal controls.Liver biopsies were collected regularly from all animals , and the Kupffer cells were isolated , purified and primarily cultured.The techniques of flow cytometry , immunohistochemistry, lysosomal fluorescent probe staining and real-time RT-PCR were applied to determine the number and function of these Kupffer cells .RESULTS: The result showed that the count and proportion of CD 163+cells in group A were significantly higher than those in group B and group C ( P<0.05).Meanwhile, the fluorescence intensity levels of lysosomal , the number of lysozyme-positive cells and the mRNA ex-pression level of TNF-αin the Kupffer cells in group A were significantly lower than those in group B and group C ( P<0.05).CONCLUSION:Kupffer cells may play a regulatory role during host’s chronic HBV infection.
5.Relationships of Inflammatory Factors and Risk Factors with Different Target Organ Damage in Essential Hypertension Patients
Lai CHUN?LIN ; Xing JIN?PING ; Liu XIAO?HONG ; Qi JIE ; Zhao JIAN?QIANG ; Ji YOU?RUI ; Yang WU?XIAO ; Yan PU?JUAN ; Luo CHUN?YAN ; Ruan LU?FANG
Chinese Medical Journal 2017;(11):1296-1302
Background: Atherosclerosis (AS) is an inflammatory disease. Inflammation was considered to play a role in the whole process of AS. This study aimed to analyze the relationships of inflammatory factors and risk factors with different target organ damages (TOD) in essential hypertension (EH) patients and to explore its clinical significance. Methods: A total of 294 EH patients were selected and divided into four groups according to their conditions of TOD. Forty?eight healthy subjects were selected as control. The clinical biochemical parameters, serum amyloid A, serum tryptase, and lipoprotein?associated phospholipase A2 (Lp?PLA2) in each group were detected, and the related risk factors were also statistically analyzed. Results: Fibrinogen (Fbg) was the most significant independent risk factor in acute coronary syndrome (ACS) group (odds ratio [OR]:22.242, 95% confidence interval [CI]: 6.458–76.609, P < 0.001) with the largest absolute value of the standardized partial regression coefficient B''(b'': 1.079). Lp?PLA2 was the most significant independent risk factor in stroke group (OR: 13.699, 95% CI: 5.236–35.837, P < 0.001) with b'' = 0.708. Uric acid (UA) was the most significant independent risk factor in renal damage group (OR: 15.307, 95%CI: 4.022–58.250, P < 0.001) with b'' = 1.026. Conclusions: Fbg, Lp?PLA2, and UA are the strongest independent risk factors toward the occurrence of ACS, ischemic stroke, and renal damage in EH patients, thus exhibiting the greatest impacts on the occurrence of ACS, ischemic stroke, and renal damage in EH patients, respectively.
6.Sperm DNA integrity of infertile males with hepatitis B virus infection.
Tian-Qin DENG ; Yong-Han HUANG ; Jin-Yi ZHEN ; Jian-Tang LU ; Ying-Chang LI ; Xing-Yin TAN ; Xiong-Chun LIU ; Jian-Bo RUAN ; Hai-Bin ZHANG
National Journal of Andrology 2013;19(1):72-76
OBJECTIVETo investigate sperm DNA integrity in male infertility patients with hepatitis B virus (HBV) infection.
METHODSThis study included 90 infertile men with HBV infection (group A), 82 infertile men without HBV infection (group B) and 70 normal fertile men (group C). We detected sperm DNA integrity among the subjects, including DNA fragmentation index (DFI) and high DNA stainability (HDS), by sperm chromatin structure assay (SCSA), and compared them among the three groups.
RESULTSDFI was higher in group A ([28.17 +/- 13.06]%) than in B ([26.64 +/- 9.79]%) and C ([15.67 +/- 4.73]%), significantly higher in A and B than in C (P < 0.05) but with no significant difference between A and B (P > 0.05). HDS was higher in group A ([10.83 +/- 5.601]%) than in B ([9.04 +/- 3.48]%) and C ([8.04-2.25]%), with significant difference between A and C (P < 0.05).
CONCLUSIONSperm DNA integrity of infertile males is significantly different from that of normal fertile men, and infertility with HBV infection further impairs sperm DNA, which is manifested by abnormal sperm nuclear maturity.
Adult ; Case-Control Studies ; Chromatin ; DNA ; genetics ; DNA Damage ; Hepatitis B ; pathology ; Hepatitis B virus ; Humans ; Infertility, Male ; genetics ; virology ; Male ; Sperm Count ; Spermatozoa ; pathology ; Young Adult
7.Liver and cardiac iron overload is harmful to HLA-identical hematopoietic stem cell transplantation in thalassemia children: an MRI detection
jing Wen YANG ; yun Jian LIAO ; yun Jian WEN ; sheng Yong RUAN ; bai Li CHEN ; lin Yue HE ; fu Chun LI ; dong Xue WU
Chinese Journal of Tissue Engineering Research 2017;21(33):5293-5298
BACKGROUND: The majority of children with β-thalassemia major have iron overload, and iron overload may have negative effects on hematopoietic stem cell transplantation. OBJECTIVE: To assess the effects of liver and cardiac iron overload detected by magnetic resonance imaging (MRI) T2* on HLA-identical allogeneic hematopoietic stem cell transplantation in children with β-thalassemia major. METHODS: Eighty-one children with β-thalassemia major who were over 3 years of age and could cooperate with MRI detection were subjected to liver and heart MRI T2* tests before or after HLA-identical allogeneic hematopoietic stem cell transplantation. According to the test results, we calculated the liver and cardiac iron content, defined as an indicator of liver and heart iron overload. Then, there was a correlation analysis between the liver and cardiac iron content and serum ferritin, time of hematopoietic reconstitution, mortality rate, implantation rate and the morbidity of transplantation related complications, such as graft-versus-host disease, infections, autoimmune hemolysis, pancytopenia, hepatic veno-occlusive disease, septicemia. RESULTS AND CONCLUSION: The liver iron content was positively correlated with the time of hemoglobin implantation (r=0.229, P=0.043), and the cardiac iron content were positively correlated with the mortality rate (r=0.266, P=0.017); the serum ferritin level was negatively correlated with the implantation rate (r=-0.289, P=0.009), and positively correlated with the morbidity of septicemia (r=0.251, P=0.024) and pancytopenia (r=0.276, P=0.013). Therefore, iron overload exerts negative effects on HLA-identical allogeneic hematopoietic stem cell transplantation in β-thalassemia major children, and it is necessary to detect serum ferritin level and assess liver and cardiac iron overload before cell transplantation.
8.The role of extracellular ATP in the male reproductive tract.
Wen-Liang ZHOU ; Wu-Lin ZUO ; Ye-Chun RUAN ; Zhe WANG ; Jian-Yang DU ; Yuan XIONG ; Hsiao-Chang CHAN
Acta Physiologica Sinica 2007;59(4):487-494
In addition to its well established role as a neurotransmitter, extracellular ATP has been considered as a paracrine/autocrine factor, either released from sperm or epithelial cells, in the male reproductive tract and shown to play a versatile role in modulating various reproductive functions. This review summarizes the signal pathways through which ATP induces anion secretion by the epithelia of the epididymis, as well as its epithelium-dependent modulation of smooth muscle contraction of the vas deferens. Finally, the overall role of ATP in coordinating various reproductive events in the male genital tract is discussed.
Adenosine Triphosphate
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physiology
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Animals
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Epididymis
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physiology
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Epithelium
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physiology
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Humans
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Male
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Muscle Contraction
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Muscle, Smooth
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physiology
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Signal Transduction
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Urogenital System
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physiology
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Vas Deferens
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physiology
9.Factors influencing long-term hepatitis B virus infection of the tree shrew (Tupaia belangeri chinensis) as an in vivo model of chronic hepatitis B.
Qi WANG ; Chun YANG ; Jian-jia SU ; Ji CAO ; Chao OU ; Fang YANG ; Jing-jing ZHANG ; Jun-lin SHI ; Dou-ping WANG ; Xiao-juan WANG ; Jia WAN ; Ping RUAN ; Yuan LI
Chinese Journal of Hepatology 2012;20(9):654-658
OBJECTIVETo determine the methods for establishing an in vivo model of long-term hepatitis B virus (HBV) infection in the Chinese tree shrew (Tupaia belangeri chinensis).
METHODSSeventy-seven neonate (1-3 days old) and 49 young adult (2 weeks to 1 year old) tree shrews were inoculated with different HBV sources (chronic hepatitis B (CHB) human patient serum, single or pooled; HBV-infected tree shrew serum, single only; HBV-infected HepG2.2.15 cells' culture medium supernatant; HBV genome-transfected HepG2.2.15 cells' culture medium supernatant) through various routes of injection (subcutaneous, intraperitoneal, and direct liver via abdominal skin; adults also received intravenous and indirect liver via spleen). Serum and liver biopsies were collected from the animals at various time points post-inoculation for detection of HBV markers by fluorescence quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, time-resolved immunofluorescence, Southern blotting, dot blotting, immunohistochemistry, and microscopy.
RESULTSAmong the neonatal group of tree shrews, six (7.8%) were confirmed as HBV-infected for more than 72 (up to 228) weeks after inoculation and another seven (9.1%) were suspected of persistent infections. None of the young adult tree shrews developed persistent infection. Inoculation with single-source serum from either CHB humans or tree shrews were responsible for the most cases of infections, and the subcutaneous injection produced more infections than the other inoculation routes. The most reliable methods of determining HBV infection status were detection of serum HBV immunoreactive markers and intrahepatic HBV DNA.
CONCLUSIONIn order to establish an in vivo model of CHB in the tree shrew, the animals should be inoculated in the neonatal period using subcutaneous injection.
Animals ; Disease Models, Animal ; Female ; Hep G2 Cells ; Hepatitis B virus ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Tupaia
10.Analysis of gag-specific IFN-γsecretion responses in HIV-1 B/C recombinant Infectors
Hong-Wei LIU ; Kun-Xue HONG ; Yuan YUAN ; Chun-Hua LIU ; Jian-Ping CHEN ; Yu-Hua RUAN ; Zhe WANG ; Yi-Ming SHAO
Chinese Journal of Experimental and Clinical Virology 2011;25(5):358-360
Objective To characterize the Gag-Specific T lymphocyte responses and identify immunodominant region recognized in Chinese HIV-1 recombinant subtype B/C infectors.Methods 10 antiretroviral treatment(ART) na?ve HIV-1 recombinant subtype B/C infectors with infected time in 1 year,25 ART-na?ve infectors with infected time > 3 years and 10 HIV-1-seronegative healthy individuals were enrolled.HIV-1-speeifie T lymphocyte responses were analyzed by an IFN-γ Elispot assay against 123 overlapping peptides spanning HIV-1 Gag protein in the present study.Results Gag-specific T lymphocyte responses of interferon-gamma secretion were identified in 8(8/10) Chinese HIV-1 recombinant subtype B/C infectors with infected time in 1 year,the specific T lymphocytes are mainly targeted at five seperated peptides.Responses were identified in 17 (68%) infectors with infected time more than 3 years,the specific T lymphocytes are mainly targeted at one peptide in p17 and six in p24.There was obviously positive correlation(P =0.0318,r =0.519 ) between the magnitude of responses and viremia in infectors infected time > 3 years.The magnitude of response in infectors infected in 1 year was significantly higher than group infected time > 3 years( P =0.021 ).None of healthy individuals produced positive responses.Conclusions HIV-1 recombinant subtype B/C Infectors at different stages of diseases recognize different region of gag.