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1.
Journal of Zhejiang University. Medical sciences ; (6): 90-96, 2021.
Article in English | WPRIM | ID: wpr-879942

ABSTRACT

To investigate the expression of xenotropic and polytropic retrovirus receptor 1 () in papillary thyroid cancer (PTC) and its clinical implication. The HPA and UALCAN databases were used to explore the expression of XPR1 in PTC and normal tissues. The cBioPortal database was used to obtain the clinical data of PTC patients and gene expression profile. The correlation of expression with gender,age,sub-types,T stage,N stage,M stage and clinical stage of patients were analyzed. Cox regression was conducted to analysis the factors affecting the prognosis of PTC patients. The mutation of was assessed through cBioPortal database. GO and KEGG analyses were used to explore the related biological pathway of involved in PTC. HPA database analysis showed that XPR1 was highly expressed in PTC tissue compared with normal tissues. UALCAN analysis displayed that expression was significantly higher in PTC tissue compared with normal tissues (0.05). Cox regression analysis showed that was an independent prognostic factor of PTC patients (=2.894,<0.05). The cBioPortal database indicated that the mutation appeared in 6% PTC patients; the mutation type mainly was missense and the mutation point was located at the E615K. Enrichment analysis indicated that might affect the PTC progression through involvement in metabolic pathway. is highly expressed in PTC tissues,which is associated with the prognosis of patients. Metabolic pathway associated with might play an important role in PTC progression,indicating that might be a novel biomarker for diagnosis and treatment of PTC.


Subject(s)
Humans , Prognosis , Receptors, G-Protein-Coupled/genetics , Receptors, Virus/genetics , Thyroid Cancer, Papillary/genetics , Thyroid Neoplasms/genetics
3.
Chinese Journal of Contemporary Pediatrics ; (12): 1344-1348, 2020.
Article in Chinese | WPRIM | ID: wpr-879800

ABSTRACT

With the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) all over the world, there is an increasing number of children with such infection. Angiotensin-converting enzyme 2 (ACE2), one of the binding sites for SARS-CoV-2 infection in humans, can bind to viral spike proteins, allowing transmembrane serine protease (TMPRSS2) to activate S-protein to trigger infection and induce the production of various inflammatory factors such as interleukin-1, interferon-l, and tumor necrosis factor. Compared with adults, children tend to have lower expression levels of ACE2 and TMPRSS2, which are presumed to be associated with milder symptoms and fewer cases in children. The article summarizes the research advances in the role of ACE2 during SARS-CoV-2 infection, in order to help understand the pathogenic mechanism of SARS-CoV-2 and provide a reference for better development of drugs and vaccines to prevent and treat coronavirus disease 2019 in children.


Subject(s)
Child , Humans , Angiotensin-Converting Enzyme 2/metabolism , COVID-19 , Receptors, Virus/metabolism , SARS-CoV-2 , Serine Endopeptidases/metabolism
4.
Journal of Zhejiang University. Medical sciences ; (6): 315-323, 2020.
Article in Chinese | WPRIM | ID: wpr-828487

ABSTRACT

OBJECTIVE@#To provide data support for the study of pathogenic mechanism of SARS-CoV-2 at the molecular level, and provide suitable candidate targets for vaccine, antibody and drug research and development through comparative analysis for structural characteristics and epitopes of S protein of SARS-CoV-2 and SARS-CoV.@*METHODS@#Based on the reference sequences of S protein, physical and chemical properties, hydrophobicity, signal peptide, transmembrane region, domain, secondary structure, tertiary structure analysis and antigenic epitopes prediction were carried out. Meanwhile, the tissue expression, related pathways and reactome pathways of angiotensis Ⅰ converting enzyme 2 (ACE2) and C-type lectin domain family 4 member M (CLEC4M) receptors were analyzed.@*RESULTS@#The amino acid sequence of S protein of SARS-CoV-2 and SARS-CoV has a 75.80% consistency. The structural characteristics of the two coronaviruses are highly consistent, but the secondary structure and tertiary structure of SARS-CoV-2 is not as obvious as SARS-CoV. ACE2 and CLEC4M are expressed in alimentary system, heart, kidney, lung and placenta. The main related the pathways of renin-angiotensin system, protein digestion and absorption pathway, and the reactome pathways of metabolism of angiotensinogen to angiotensins, GPCR ligand binding, are related to typical symptoms of coronavirus disease 2019 induced by SARS-CoV-2. Three pairs of highly or completely homologous epitopes of S protein were obtained. The 600-605, 695-703 and 888-896 amino acid residues in SARS-CoV-2 were highly homologous with 586-591, 677-685 and 870-878 amino acid residues in SARS-CoV, respectively.@*CONCLUSIONS@#The similarity of S protein of SARS-CoV-2 and SARS-CoV determines that they have similar infection patterns and clinical manifestations. The candidate epitopes with high reliability can provide reference for virus diagnosis and vaccine development.


Subject(s)
Humans , Betacoronavirus , Cell Adhesion Molecules , Coronavirus Infections , Epitopes , Lectins, C-Type , Ligands , Pandemics , Peptidyl-Dipeptidase A , Pneumonia, Viral , Receptors, Cell Surface , Receptors, Virus , Reproducibility of Results , Spike Glycoprotein, Coronavirus
5.
Protein & Cell ; (12): 590-600, 2017.
Article in English | WPRIM | ID: wpr-756983

ABSTRACT

Entero virus 71 (EV71) causes hand, foot, and mouth disease (HFMD) and occasionally leads to severe neurological complications and even death. Scavenger receptor class B member 2 (SCARB2) is a functional receptor for EV71, that mediates viral attachment, internalization, and uncoating. However, the exact binding site of EV71 on SCARB2 is unknown. In this study, we generated a monoclonal antibody (mAb) that binds to human but not mouse SCARB2. It is named JL2, and it can effectively inhibit EV71 infection of target cells. Using a set of chimeras of human and mouse SCARB2, we identified that the region containing residues 77-113 of human SCARB2 contributes significantly to JL2 binding. The structure of the SCARB2-JL2 complex revealed that JL2 binds to the apical region of SCARB2 involving α-helices 2, 5, and 14. Our results provide new insights into the potential binding sites for EV71 on SCARB2 and the molecular mechanism of EV71 entry.


Subject(s)
Animals , Humans , Mice , Amino Acid Sequence , Antibodies, Monoclonal , Chemistry , Genetics , Metabolism , Binding Sites , Cell Line , Crystallography, X-Ray , Enterovirus A, Human , Genetics , Allergy and Immunology , Fibroblasts , Virology , Gene Expression , HEK293 Cells , Immunoglobulin Fab Fragments , Chemistry , Genetics , Metabolism , Lysosome-Associated Membrane Glycoproteins , Chemistry , Genetics , Allergy and Immunology , Models, Molecular , Protein Binding , Protein Conformation, alpha-Helical , Protein Conformation, beta-Strand , Protein Interaction Domains and Motifs , Receptors, Scavenger , Chemistry , Genetics , Allergy and Immunology , Receptors, Virus , Chemistry , Genetics , Allergy and Immunology , Recombinant Fusion Proteins , Chemistry , Genetics , Allergy and Immunology , Sequence Alignment , Sequence Homology, Amino Acid , Sf9 Cells , Spodoptera , Thermodynamics
6.
Chinese Journal of Contemporary Pediatrics ; (12): 24-28, 2016.
Article in Chinese | WPRIM | ID: wpr-279903

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the values of urinary netrin-1 and kidney injury molecule-1 (KIM-1) in the early diagnosis of acute kidney injury (AKI) induced by neonatal asphyxia.</p><p><b>METHODS</b>A total of 80 full-term neonates with asphyxia were enrolled (mild asphyxia: 34 neonates; severe asphyxia: 46 neonates). Forty normal full-term neonates were selected as the control group. Urinary samples were collected from the neonates in the three groups within 12 hours and 13-48 hours after birth. ELISA was applied to measure urinary levels of netrin-1 and KIM-1. Peripheral venous blood samples were also collected to measure serum creatinine (Scr) level.</p><p><b>RESULTS</b>Compared with the control group, the asphyxia group had significantly higher urinary levels of netrin-1 and KIM-1 within 48 hours after birth and a significantly higher Scr level within 13-48 hours after birth (P<0.05). The neonates in the AKI group had significantly higher urinary levels of netrin-1 and KIM-1 and Scr level within 48 hours after birth than those in the non-AKI group (P<0.05). The areas under the receiver operating characteristic curve for urinary netrin-1 and KIM-1 levels within 12 hours after birth to predict AKI after asphyxia were 0.878 (95% CI: 0.775-0.981; P<0.01) and 0.899 (95% CI: 0.829-0.969; P<0.01), respectively. Any two indicators of urinary netrin-1 level, urinary KIM-1 level, and Scr level within 12 hours after neonatal asphyxia had a positive correlation (P<0.05).</p><p><b>CONCLUSIONS</b>Urinary netrin-1 and KIM-1 levels increase significantly when neonates with asphyxia develop AKI. Urinary netrin-1 and KIM-1 can be used as indicators for the early diagnosis of AKI after asphyxia.</p>


Subject(s)
Female , Humans , Infant, Newborn , Male , Acute Kidney Injury , Diagnosis , Urine , Asphyxia Neonatorum , Hepatitis A Virus Cellular Receptor 1 , Membrane Glycoproteins , Urine , Nerve Growth Factors , Urine , Netrin-1 , Receptors, Virus , Tumor Suppressor Proteins , Urine
7.
Journal of Forensic Medicine ; (6): 97-104, 2015.
Article in Chinese | WPRIM | ID: wpr-983971

ABSTRACT

OBJECTIVE@#To understand the correlation of enterovirus 71 (EV71), P-selectin glycoprotein ligand-1 (PSGL-1), and scavenger receptor B2 (SCARB2) and to explore the possible pathway and mechanism of EV71 infection by observing the expression of EV71, PSGL-1 and SCARB2 in tissues of infants with brain stem encephalitis.@*METHODS@#The organs and tissues of infants with EV71-VP1 positivity in their brain stems were chosen. Expression and distribution of EV71-VP1, PSGL-1, and SCARB2 were detected and compared by immunohistochemistry.@*RESULTS@#Strong staining of EV71 -VP1 was observed in the neuron, glial cells, the inflammatory cells of perivascular cuffing, parietal cells of the gastric fundus gland while alveolar macrophages, intestinal gland epithelium cells, mucosa lymphoid nodule and lymphocyte of palatine tonsil showed moderate staining and weak staining were displayed in mesenteric lymph nodes and lymphocyte of spleen. PSGL-1 expression was detected in parietal cells of the gastric fundus gland, tonsillar crypt squamous epithelium, alveolar macrophages and leukocytes in each tissue. SCARB2 expression was observed in all the above tissues except the intestines and spleen.@*CONCLUSION@#The distribution of EV71 correlates with SCARB2 expression. SCARB2 plays an important role in virus infection and replication. Stomach may be an important site for EV71 replication.


Subject(s)
Humans , Infant , Brain Stem/virology , Encephalitis, Viral/virology , Enterovirus A, Human/metabolism , Enterovirus Infections/virology , Immunohistochemistry , Leukocytes , Lysosome-Associated Membrane Glycoproteins , Membrane Glycoproteins/metabolism , Receptors, Scavenger/metabolism , Receptors, Virus/metabolism
8.
Journal of Central South University(Medical Sciences) ; (12): 1083-1088, 2015.
Article in Chinese | WPRIM | ID: wpr-815222

ABSTRACT

OBJECTIVE@#To evaluate the early predictive and diagnostic significance of the acute kidney injury (AKI) associated biomarkers for patients in the intensive care unit (ICU).
@*METHODS@#From January to June, 2014, relevant clinical data of participants were collected upon admission to the intensive care unit (ICU) in Affiliated Hospital of Zunyi Medical College. Levels of serum cystatin C (sCys C), neutrophil gelatinase-associated lipocalin (sNGAL), urinary neutrophil gelatinase-associated lipocalin (uNGAL), urinary kidney injury molecule-1 (uKIM-1), interleukin-18 (uIL-18), and N-acetyl-beta-D-glucosaminidase (uNAG) were detected by enzyme linked immune sorbent assay (ELISA), and compared between AKI and non-AKI patients. Diagnostic significance of these biomarkers was evaluated by a receiver operating characteristic (ROC) curve and the area under the ROC curve.
@*RESULTS@#A total of 176 patients were enrolled in this study. Among them, 71 patients were diagnosed as AKI, in which 57 patients hospitalized with AKI and 14 developed AKI after 24 h hospitalization. The renal replacement therapy ratio was increased with the progress of clinical stage for AKI. AKI mortality rate was 18.8% (46.5% of the total number of deaths). The levels of sCys C, sNGAL, uNGAL, and uIL-18 in AKI patients were increased compared with those in the non-AKI patients (P<0.05). With the progress of AKI, sCys C, and uNGAL levels were also elevated. In 14 patients who suffered from AKI 24 h after hospitalization, the average levels of sCys C, uNGAL, uIL-18, and uKIM-1 were significantly increased (P<0.05). Sensitivity and specificity of the uNGAL, sCys C, and uIL-18 in AKI diagnosis were 97.2%, 76.1%, 54.9% and 93.3 %, 96.2%, 78.1%, respectively. The areas under the ROC curve of uNGAL, sCys C, and uIL-18 were 0.99, 0.90, and 0.69, respectively.
@*CONCLUSION@#uNGAL, sCys C and uIL-18 can be used to predict and diagnose AKI, and to evaluate the AKI clinical stage.


Subject(s)
Humans , Acetylglucosaminidase , Urine , Acute Kidney Injury , Blood , Diagnosis , Urine , Acute-Phase Proteins , Urine , Biomarkers , Blood , Urine , Case-Control Studies , Cystatin C , Blood , Enzyme-Linked Immunosorbent Assay , Hepatitis A Virus Cellular Receptor 1 , Intensive Care Units , Interleukin-18 , Urine , Lipocalin-2 , Lipocalins , Blood , Urine , Membrane Glycoproteins , Urine , Proto-Oncogene Proteins , Blood , Urine , ROC Curve , Receptors, Virus , Sensitivity and Specificity
9.
Journal of Central South University(Medical Sciences) ; (12): 276-280, 2015.
Article in Chinese | WPRIM | ID: wpr-815172

ABSTRACT

OBJECTIVE@#To detect the levels of neutrophil gelatinase-associated lipocalin (NGAL), cystatin C (Cys-C ) in blood and the level of kidney injury molecule 1 (KIM-1) in urine in elderly patients with renal calculi at diff erent times, and to explore the eff ect of percutaneous nephrostolithotomy (PCNL) combined with flexible ureteroscopy (FU) on early postoperative renal function.@*METHODS@#A total of 46 patients with renal calculi were selected, and their blood or urine specimens were collected respectively at preoperative and postoperative 2, 12, 24, 48, and 72 h. The concentrations of NGAL, Cys-C, KIM-1 were detected.@*RESULTS@#The levels of NGAL and Cys-C began to increase respectively at postoperative 2 and 12 h, and reached peak at postoperative 12 to 24 h. There was significant difference in the levels of NGAL and Cys-C between the postoperative 12 and 2 h or between postoperative 48 and 24 h (all P<0.05). The levels of NGAL and Cys-C began to decline and eventually returned to preoperative levels respectively at postoperative 48 and postoperative 72 h. The KIM-1 began to increase at postoperative 2 h and peaked at postoperative 24 h, which was significant difference between the postoperative 24 and 12 h or postoperative 48 and 24 h (both P<0.05). The level of KIM-1 began to decline and eventually returned to preoperative levels at postoperative 48 h.@*CONCLUSION@#After the combined treatment of percutaneous nephrostolithotomy with flexible ureteroscopy, the concentrations of NGAL, Cys-C and KIM-1 are significantly increased, suggesting injuries on renal function. The time of renal tubular injury and recovery is earlier than that of renal glomerulus.


Subject(s)
Aged , Humans , Acute-Phase Proteins , Urine , Cystatin C , Blood , Urine , Hepatitis A Virus Cellular Receptor 1 , Kidney , Kidney Calculi , General Surgery , Lipocalin-2 , Lipocalins , Blood , Urine , Membrane Glycoproteins , Blood , Urine , Nephrostomy, Percutaneous , Postoperative Period , Proto-Oncogene Proteins , Blood , Urine , Receptors, Virus , Blood , Ureteroscopy
10.
Chinese Journal of Virology ; (6): 97-105, 2015.
Article in Chinese | WPRIM | ID: wpr-280288

ABSTRACT

Hepatitis c virus (HCV) infection has become one of the global public health problem,while there is no vaccine to prevent HCV infection, the so-called "cocktail" therapy that use a combination of drugs targeting multiple steps in the HCV infection cycle could achieve better curative effect. the process of HCV entering into host cell is the important step of drug intervention, in which HCV envelope protein El and E2, Host cell factors including Heparan sulfate(HS), CD81, scavenger receptor class B type I (SR-BI), Occludin (OCLD), Claudin (CLDN), low densitity lipoprotein receptor (LDLR), dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN), Liver/lymph node specific ICAM-3-grabbing integrin(L-SIGN), trans- ferrin receptor 1 (TfR1) and so on play a important role. The virus and the host factors can be used as targets of hcv entry inhibitors many studies have shown that as novel and promising compounds, HCV entry inhibitors combinating with other drugs can be more effective in the treatment of HCV, this paper have re- viewed targets and inhibitors of HCV enterring into host cell since 1990s.


Subject(s)
Animals , Humans , Antiviral Agents , Pharmacology , Hepacivirus , Physiology , Hepatitis C , Genetics , Metabolism , Virology , Receptors, Virus , Genetics , Metabolism , Viral Envelope Proteins , Genetics , Metabolism , Virus Internalization
11.
Chinese Journal of Virology ; (6): 164-169, 2015.
Article in Chinese | WPRIM | ID: wpr-280279

ABSTRACT

Interactions between noroviruses (NoVs) and the receptors of histo-blood group antigens (HB-GAs) affect the infectivity and host susceptibility of NoVs. We elucidated the binding profile of a GII. 12 NoV to HBGAs. First, we synthesized the P domain sequence of the GII. 12 NoV strain Pune (GenBank accession number EU921353). Protein of the P domain was expressed in a prokaryotic system. Formation of the P particle was monitored by gel-filtration chromatography. Antiserum was prepared by immunization of mice with GII. 12 P particles. The binding profile of the GII. 12 NoV Pune strain was determined by binding of the P particle with a panel of saliva samples with various known HBGAs phenotypes. The GII. 12 NoV was bound strongly to saliva samples of subjects with B and AB types and weakly to A, O secretor, and non-secretor saliva samples, suggesting higher affinity with B antigen by GII. 12 NoV. These results were consistent with those determined by a previous crystallography study of GII. 12 NoV. These data suggested that individuals with B and AB blood types may be more susceptible to infection by GII. 12 NoV compared with those with other blood types. Our findings may provide a basis for the prevention and control of an epidemic of GII. 12 NoV.


Subject(s)
Animals , Female , Humans , Mice , Blood Group Antigens , Metabolism , Caliciviridae Infections , Metabolism , Virology , Gastroenteritis , Metabolism , Virology , Genotype , Mice, Inbred BALB C , Norovirus , Genetics , Metabolism , Protein Binding , Receptors, Virus , Metabolism , Viral Proteins , Genetics , Metabolism
12.
Protein & Cell ; (12): 814-824, 2015.
Article in English | WPRIM | ID: wpr-757174

ABSTRACT

Ebolavirus can cause hemorrhagic fever in humans with a mortality rate of 50%-90%. Currently, no approved vaccines and antiviral therapies are available. Human TIM1 is considered as an attachment factor for EBOV, enhancing viral infection through interaction with PS located on the viral envelope. However, reasons underlying the preferable usage of hTIM-1, but not other PS binding receptors by filovirus, remain unknown. We firstly demonstrated a direct interaction between hTIM-1 and EBOV GP in vitro and determined the crystal structures of the Ig V domains of hTIM-1 and hTIM-4. The binding region in hTIM-1 to EBOV GP was mapped by chimeras and mutation assays, which were designed based on structural analysis. Pseudovirion infection assays performed using hTIM-1 and its homologs as well as point mutants verified the location of the GP binding site and the importance of EBOV GP-hTIM-1 interaction in EBOV cellular entry.


Subject(s)
Humans , Ebolavirus , Metabolism , Flow Cytometry , Glycoproteins , Metabolism , Hepatitis A Virus Cellular Receptor 1 , Hepatitis A Virus Cellular Receptor 2 , Membrane Glycoproteins , Metabolism , Membrane Proteins , Metabolism , Protein Binding , Receptors, Virus , Metabolism , Surface Plasmon Resonance , Viral Envelope Proteins , Metabolism , Viral Proteins , Metabolism
13.
Chinese Journal of Cardiology ; (12): 301-304, 2014.
Article in Chinese | WPRIM | ID: wpr-316469

ABSTRACT

<p><b>OBJECTIVE</b>To explore the predict value of monitoring changes of urinary neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1(KIM-1) after coronary angiography (CAG) and percutaneous coronary intervention (PCI) on the early diagnosis of contrast-induced nephropathy(CIN).</p><p><b>METHODS</b>One hundred and sixty patients underwent CAG and PCI were enrolled in this prospective study. There were 14 patients with CIN and non-CIN patients were selected with the proportion of 2: 1 (n = 28).Serum creatinine (SCr) was measured before and at 24, 48 and 72 h after the procedure. Urinary NGAL and KIM-1 were measured before and at 4 and 24 h after the procedure. The relationship between NGAL, KIM-1 and CIN were analyzed. Receiver operating characteristic (ROC) curve and area under the ROC curve (AUC) were used to analyze the diagnostic sensitivity and specificity of CIN by urinary NGAL and KIM-1.</p><p><b>RESULTS</b>(1) The values of urinary NGAL was significantly higher in the CIN group than in non-CIN group at 4 h after CAG or PCI (P < 0.01); the value of urinary NGAL was significantly increased from the baseline to 4 h after the procedure in the CIN group (P < 0.01). (2) Uurinary KIM-1 levels of CIN group was significantly higher than in non-CIN group at 24 h after the CAG or PCI (P < 0.01) ; the urinary KIM-1 levels was significantly increased from baseline to 24 h after the procedure in the CIN group (P < 0.01). (3) Pearson correlation analysis showed that there was a positive correlation between urinary NGAL and SCr (r = 0.814, P < 0.01) and urinary KIM-1(r = 0.758, P < 0.01) in the CIN group. (4) ROC curve analysis showed that the AUC for urinary NGAL was 0.897. When the cut-off value of NGAL was set at 11.950 µg/L, the sensitivity and specificity for the diagnosis of CIN were 92.9% and 71.4%, respectively. The AUC for urinary KIM-1 was 0.839. With the cut-off value of urinary KIM-1 set as 4.595 µg/L, the diagnostic sensitivity and specificity for CIN were 85.7% and 71.4%, respectively.</p><p><b>CONCLUSIONS</b>Urinary NGAL serves as a good biomarker for early diagnosis of CIN suggesting acute kidney injury at 4 h post CAG and PCI. Urinary KIM-1 can reflect the change of renal function after contrast injection earlier than SCr and may also be a good biomarker for early diagnosis of CIN.</p>


Subject(s)
Aged , Humans , Middle Aged , Acute-Phase Proteins , Urine , Contrast Media , Coronary Angiography , Hepatitis A Virus Cellular Receptor 1 , Kidney Diseases , Lipocalin-2 , Lipocalins , Urine , Membrane Glycoproteins , Urine , Percutaneous Coronary Intervention , Predictive Value of Tests , Prospective Studies , Proto-Oncogene Proteins , Urine , Receptors, Virus , Sensitivity and Specificity
14.
Chinese Journal of Virology ; (6): 303-309, 2014.
Article in Chinese | WPRIM | ID: wpr-356599

ABSTRACT

Rotaviruses, which are recognized as one of the major etiological agents among infants and young children with diarrhea, consist of three concentric layers of protein capsid with the enclosed double-stranded RNA genome. Rotaviruses infect host cells mainly by identifying the specific receptors on cell surfaces and binding to them. Therefore, receptors are important factors for viruses infecting cells. So far, there have been many receptors found to be involved in rotavirus infection, including sialic acid, integrin, Toll-like receptor, and blood group antigen. This article provides an overview of receptors involved in rotavirus infection.


Subject(s)
Animals , Humans , Receptors, Virus , Genetics , Metabolism , Rotavirus , Genetics , Physiology , Rotavirus Infections , Genetics , Metabolism , Virology
15.
Biomedical and Environmental Sciences ; (12): 641-645, 2014.
Article in English | WPRIM | ID: wpr-270556

ABSTRACT

Melamine in combination with cyanuric acid has been considered to be more toxic than either melamine or cyanuric acid alone. The objective of this study was designed to evaluate the combined genotoxicity and cytotoxicity of melamine (M) and cyanuric acid (C) at three mass ratios (1:1, 1:2, 2:1). MC (1:1), MC (1:2), and MC (2:1) were evaluated for their potential genotoxic risk, at gene level by Ames test, and at chromosomal level by micronucleus test. In order to evaluate cytotoxicity in HEK-293 cells, the MTT assay was included. Western blot was also employed to investigate the renal injury molecule-1 (Kim-1) expression in HEK-293 cells exposed to MC. Neither genotoxicity at gene level nor at chromosomal level was observed for MC (1:1), MC (1:2), and MC (2:1). Based on MTT assay, three ratios of MC at 82.5 and 165 µg/mL slightly inhibited viability of HEK-293 cells (P<0.05). MC (1:1) at 41.25 and 82.50 µg/mL could elevate the Kim-1 expression in HEK-293 cells.


Subject(s)
Humans , Cell Survival , HEK293 Cells , Hepatitis A Virus Cellular Receptor 1 , Membrane Glycoproteins , Metabolism , Receptors, Virus , Metabolism , Triazines , Pharmacology
16.
Chinese Journal of Virology ; (6): 106-111, 2013.
Article in Chinese | WPRIM | ID: wpr-339967

ABSTRACT

The receptor-binding domain(RBD) protein of HCoV-NL63 is a major target in the development of diagnostic assay and vaccine, it has a pivotal role in receptor attachment, viral entry and membrane fusion. In this study, we prepared 2 purified recombinant HCoV-NL63 RBD proteins using in E. coli system and identified the proteins by Western blotting. We first optimized codon and synthesized the RL (232-684aa)coding gene, then amplified the RL or RS(476-616aa) coding gene via PCR using different primers . The RL or RS coding gene was cloned into the pM48 expression vector fused with TrxA tag. The RBD (RL and RS) of HCoV-NL63 were expressed majorly as inclusion body when expressed in E. coli BL21pLys S under different conditions. The expressed products were purified by affinity chromatography then analyzed by SDS-PAGE and Western blotting. Our results showed that the recombinant RBD proteins were maximally expressed at 37 degrees C with 0. 8mM IPTG induction for 4h. RL or RS protein with 95 % purity was obtained and reacted positively with anti-sera from mice immunized with the recombinant vaccinia virus (Tiantan strain) in which HCoV-NL63 RL or RS protein was expressed. In conclusion, the purified recombinant RBD proteins(RL and RS)derived from E. coli were first prepared in China and they might provide a basis for further exploring biological role and vaccine development of HCoV-NL63.


Subject(s)
Animals , Humans , Mice , Coronavirus Infections , Metabolism , Virology , Coronavirus NL63, Human , Chemistry , Genetics , Metabolism , Escherichia coli , Genetics , Metabolism , Gene Expression , Mice, Inbred BALB C , Protein Engineering , Protein Structure, Tertiary , Receptors, Virus , Metabolism , Viral Envelope Proteins , Chemistry , Genetics , Metabolism
17.
Chinese Journal of Hematology ; (12): 614-617, 2013.
Article in Chinese | WPRIM | ID: wpr-272155

ABSTRACT

<p><b>OBJECTIVE</b>To explore the expression and clinical significance of T cell immunoglobulin mucin (TIM)-1, TIM-3 and T cell-specific transcription factors T-bet and GATA-3 in spleen mononuclear cells in patients with primary immune thrombocytopenia (ITP).</p><p><b>METHODS</b>The spleen samples were obtained from 17 active ITP patients and 10 controls with spleen traumatic rupture. By using real-time quantitative polymerase chain reaction, the mRNA expressions of TIM-3, TIM1, T-bet and GATA-3 were studied in all subjects.</p><p><b>RESULTS</b>TIM-3 mRNA levels of active ITP patients were significantly decreased to (29 ± 16)% of that of control, TIM-1 mRNA levels of active ITP patients increased to (3.20 ± 2.18) folds of that of control, but the difference was not significant. The ratio of TIM-1/ TIM-3 was elevated in active ITP patients. T-bet mRNA levels were up-regulated in ITP patients by (2.82 ± 1.57) folds (P<0.05) and the expression of GATA3 was decreased by 14% folds (P<0.05) compared to controls. The ratio of T-bet/GATA3 were significantly elevated in ITP patients.</p><p><b>CONCLUSION</b>The imbalance between TIM-3 and TIM-1 expression might play an important role in pathogenesis of ITP.</p>


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Case-Control Studies , Flow Cytometry , GATA3 Transcription Factor , Metabolism , Hepatitis A Virus Cellular Receptor 1 , Hepatitis A Virus Cellular Receptor 2 , Membrane Glycoproteins , Metabolism , Membrane Proteins , Metabolism , Purpura, Thrombocytopenic, Idiopathic , Allergy and Immunology , Metabolism , RNA, Messenger , Genetics , Receptors, Virus , Metabolism , Spleen , Metabolism , Th1 Cells , Allergy and Immunology , Th2 Cells , Allergy and Immunology
18.
Chinese Journal of Contemporary Pediatrics ; (12): 302-307, 2013.
Article in Chinese | WPRIM | ID: wpr-236815

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of down-regulating Tim-3 gene in the peripheral blood mononuclear cells (PBMCs) of an asthmatic mouse model by short hairpin RNA (shRNA) and to explore the effect of Tim-3 on Th1 and Th17 cell differentiation.</p><p><b>METHODS</b>An asthmatic murine model was established by ovalbumin sensitization and challenge. PBMCs were isolated from asthmatic mice and transfected by shRNA targeting Tim-3 gene. The mRNA and protein expressions of Tim-3 were detected by quantitative PCR and Western blot. Flow cytometry analysis was performed to determine the levels of Th1 and Th17, and ELISA was performed to determine concentrations of IFN-γ, IL-4 and IL-17 in the supernatant.</p><p><b>RESULTS</b>Tim-3 mRNA expression in PBMCs was significantly increased in asthmatic mice. The mRNA and protein expression of Tim-3 decreased significantly in the shRNA group. Compared with the negative groups, Th1 cell levels increased and Th17 cell levels decreased significantly in the asthmatic groups after Tim-3 shRNA interference. In the Tim-3 shRNA interference groups concentrations of IFN-γ increased significantly while IL-17 decreased significantly.</p><p><b>CONCLUSIONS</b>Specific Tim-3 shRNA effectively silences the expression of Tim-3 and change in Tim-3 expression could affect T cell differentiation.</p>


Subject(s)
Animals , Female , Mice , Asthma , Allergy and Immunology , Therapeutics , Cell Differentiation , Gene Silencing , Hepatitis A Virus Cellular Receptor 2 , Mice, Inbred BALB C , RNA, Small Interfering , Genetics , Receptors, Virus , Genetics , Th1 Cells , Cell Biology , Th17 Cells , Cell Biology
19.
Chinese Journal of Biotechnology ; (12): 1-9, 2013.
Article in Chinese | WPRIM | ID: wpr-233275

ABSTRACT

Measles virus is an enveloped virus with a non-segmented negative-sense RNA genome. Two envelope glycoproteins on the viral surface, namely hemagglutinin (H) and membrane fusion protein (F), are responsible for the virus entry into susceptible host cells. The specific interaction between H and its cellular receptors is a key step in successful virus infection, determining the infectivity and tissue tropism of the measles virus. Thus far, three H receptors have been identified, including the complement regulatory molecule CD46, the signaling lymphocyte activation molecule (SLAM) and the cell adhesion molecule Nectin-4. Here, we reviewed our molecular understanding on the recognition mechanism of these receptors by the viral H protein, aiming to promote future studies on antiviral drug design and measles virus-based oncolytic therapy.


Subject(s)
Animals , Humans , Antigens, CD , Metabolism , Cell Adhesion Molecules , Metabolism , Hemagglutinins, Viral , Metabolism , Measles virus , Virulence , Physiology , Membrane Cofactor Protein , Metabolism , Membrane Fusion , Membrane Fusion Proteins , Metabolism , Receptors, Cell Surface , Metabolism , Receptors, Virus , Metabolism , Signaling Lymphocytic Activation Molecule Family Member 1
20.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 479-484, 2013.
Article in English | WPRIM | ID: wpr-251445

ABSTRACT

This study investigated the expression of interleukin-17 (IL-17) and T cell immunoglobulin mucin and domain-containing molecule-3 (Tim-3) in bronchoalveolar lavage fluid (BALF) of asthmatic mice and the effect of dexamethasone (DEX) on these factors. Thirty-six mice were randomly divided into three groups: normal group, asthmatic group and DEX group. The mouse model of asthma was established by sensitization with ovalbumin in both the asthmatic and DEX groups. The levels of IL-6, IL-10, IL-17 and TGF-β were measured in BALF by enzyme-linked immunesorbent assay (ELISA). The mRNA expression level of Tim-3 was detected by reverse transcription polymerase chain reaction (RT-PCR). The ratio of Tim-3+CD4+ cells to total CD4+ cells in BALF was determined by flow cytometry. Differential inflammatory cells in BALF were detected. The correlations among IL-17, IL-6, IL-10, Tim-3 and inflammatory cells were analyzed. The results showed that the levels of IL-17, IL-6 and Tim-3 were substantially increased and the IL-10 level decreased in BALF in the asthmatic mice, which was significantly reversed by DEX treatment. IL-17 expression was positively correlated with IL-6 and Tim-3 expression and the number of inflammatory cells but negatively with IL-10 expression. These results indicate that the increased expression of IL-17 and Tim-3 in BALF may be implicated in the occurrence and development of asthmatic inflammation; the mechanism by which DEX suppresses asthmatic airway inflammation involves down-regulation of IL-17 and Tim-3 levels.


Subject(s)
Animals , Female , Mice , Asthma , Drug Therapy , Genetics , Metabolism , Bronchoalveolar Lavage Fluid , Chemistry , Dexamethasone , Pharmacology , Gene Expression , Genetics , Hepatitis A Virus Cellular Receptor 2 , Interleukin-17 , Genetics , Metabolism , Mice, Inbred BALB C , Receptors, Virus , Genetics , Metabolism
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