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1.
Journal of Experimental Hematology ; (6): 1465-1470, 2018.
Article in Chinese | WPRIM | ID: wpr-689912

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the oxidative stress status and its effects on hepcidin in patients with hemoglobin H Constant Spring disease (HbH-CS).</p><p><b>METHODS</b>A total of 35 patients were enrolled in the study, including 15 splenectomized cases and 20 non-splenectomized cases. 20 healthy volunteers were selected as controls. Serum superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), oxidized glutathione (GSSG) levels, erythropoietin (EPO), serum free transferrin receptor (sFTR), growth differentiation factor 15 (GDF15) as well as the level of hepcidin were detected. Correlation analysis and multiple factor regression analysis were performed to investigate the factors affecting the iron metabolism and erythropoiesis.</p><p><b>RESULTS</b>Compared with healthy control, the SOD and GSH levels in patients with HbHCS decreased, while MDA and GSSG levels increased. The levels of SOD, MDA, GSG and GSSG were not significantly different between the patients with splenectomy and those without splenectomy. Correlation analysis showed that inpatients with HbHCS, EPO, sFTR and GDF15 correlated negatively with SOD level and positively with MDA level. EPO and sFTR levels negatively correlated with Hepcidin.</p><p><b>CONCLUSION</b>Excessive oxidative stress is present in patients with HbHCS, and hepcidin is inhibited by the upregulation of EPO and sFTR, and hence involved in iron overload in patients.</p>

2.
Chinese Journal of Contemporary Pediatrics ; (12): 49-52, 2015.
Article in Chinese | WPRIM | ID: wpr-289471

ABSTRACT

<p><b>OBJECTRIVE</b>To compare the differences in risk factors for low birth weight (LBW) between Han and Uygur full-term infants and to provide a basis for the prevention of LBW in newborn infants.</p><p><b>METHODS</b>Eighty-seven full-term LBW infants (38 Hans and 49 Uygurs) between March 2013 and June 2014 were selected as the case group, and 186 full-term normal birth weight infants (92 Hans and 94 Uygurs) were selected as the control group. A questionnaire survey was performed to investigate the related factors for LBW. Multivariate logistic regression analysis was carried out to determine the risk factors for LBW.</p><p><b>RESULTS</b>The birth weights in Uyghur LBW infants were lower than in Han ones (P<0.05). Multivariate logistic regression analysis showed that drinking (OR=2.472, P=0.015) and smoking (OR=2.323, P=0.007) by the father, pregnancy complications (OR=14.377, P<0.001), and times of pregnancy (OR=2.995, P=0.001) were the risk factors for LBW in Han infants, while drinking by the father (OR=1.968, P=0.007), times of pregnancy (OR=1.953, P=0.005), pregnancy complications (OR=10.283, P=0.002), and poor indoor environment (OR=1.367, P=0.027) were the risk factors for LBW in Uyghur infants.</p><p><b>CONCLUSIONS</b>There are differences in physical growth between Han and Uygur LBW infants. Han and Uygur infants share the same traditional risk factors for LBW, such as father's harmful behaviors like drinking, times of pregnancy, and pregnancy complications, however, the indoor environment also plays a role in the occurrence of LBW in Uygur infants.</p>


Subject(s)
Female , Humans , Infant, Newborn , Pregnancy , China , Ethnology , Infant, Low Birth Weight , Logistic Models , Pregnancy Complications , Risk Factors
3.
Chinese Journal of Virology ; (6): 636-644, 2014.
Article in Chinese | WPRIM | ID: wpr-280315

ABSTRACT

Hepatitis B virus (HBV) is a major cause of chronic liver disease, and frequently results in hepatitis, cirrhosis, and ultimately hepatocellular carcinoma. HBV polymerase (Pol) is an essential viral protein that is important for HBV replication and might be involved in the development of hepatocellular carcinoma. Protein-protein interactions appears to be crucial for its role. The aim of this study was to screen and identify the proteins that interact with Pol using a co-immunoprecipitation-based LC-MS/MS identification technique. The HBV Pol gene was amplified by polymerase chain reaction (PCR) and cloned into pCDNA3.1(+). The recombinant plasmid pCDNA3. 1(+)-Pol-flag was transfected into HeLa cells. Liquid chromatography and tandem mass spectrometry (LC-MS/MS) identified 45 proteins that co-immunoprecipitated with flag-tagged HBV Pol. Eleven of these have previously been reported as proteins that interact with HBV Pol. A proof-of-concept-based Ingenuity Pathway Analysis (IPA, www.ingenuity.com) was used to characterize the functions and pathways of these 45 identified proteins and HBV Pol. Among these proteins, four proteins may play a role in three major molecular cellular networks, and are therefore worthy of further investigation.


Subject(s)
Humans , Cell Line, Tumor , Chromatography, Liquid , Methods , Gene Products, pol , Chemistry , Genetics , Metabolism , Hepatitis B , Genetics , Metabolism , Virology , Hepatitis B virus , Chemistry , Genetics , Immunoprecipitation , Methods , Protein Interaction Maps , Software , Tandem Mass Spectrometry , Methods
4.
Chinese Journal of Experimental and Clinical Virology ; (6): 57-59, 2011.
Article in Chinese | WPRIM | ID: wpr-231193

ABSTRACT

<p><b>OBJECTIVE</b>To explore relevant between human cytomegalovirus (HCMV) infection and college students' neurobehaviors.</p><p><b>METHODS</b>87 college students were enlisted. They were tested with Bole. Neurobehavioral evaluation system (B. NES), and HCMV IgG antibody was detected after separation of serum. We analyzed the test results of B. NES by SPSS software.</p><p><b>RESULTS</b>76 college students were infected by HCMV in the past and 11 college students were not infected. The infected group scored 8.89 +/- 6.60 in depression aspect of emotion state test, while control group got 15.73 +/- 9.00. There was Significant difference between infection group and control (P < 0.05). There were no significant differences in other aspects of emotion states, study and memory, perception and mental movement (P > 0.05).</p><p><b>CONCLUSION</b>HCMV infection is associated with depression status.</p>


Subject(s)
Adult , Female , Humans , Male , Cytomegalovirus Infections , Psychology , Emotions , Learning , Memory , Students , Psychology , Universities
5.
Journal of Southern Medical University ; (12): 686-689, 2011.
Article in Chinese | WPRIM | ID: wpr-332574

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the function of hepatitis B virus polymerase (HBV Pol) in the viral life cycle by screening the proteins interacting with HBV polymerase.</p><p><b>METHODS</b>The HBV Pol gene was constructed into the pGBKT7 vector. GAL4 yeast two-hybrid system was used to screen the human liver cDNA library to obtain proteins which interacted with HBV Pol. GST-pull down assay was applied to confirm the protein interactions.</p><p><b>RESULTS</b>Ubiquitously expressed transcript (UXT) was selected by the yeast two-hybrid system. GST-pull down assay confirmed the in vitro interaction between HBV Pol and UXT.</p><p><b>CONCLUSIONS</b>UXT is a potential interactor of HBV Pol, and this protein interaction may provide clues of the function of HBV Pol in HBV life cycle.</p>


Subject(s)
Humans , Gene Products, pol , Metabolism , Hepatitis B virus , Neoplasm Proteins , Metabolism , Protein Interaction Mapping , Two-Hybrid System Techniques , Virus Replication
6.
Chinese Journal of Medical Genetics ; (6): 62-65, 2009.
Article in Chinese | WPRIM | ID: wpr-287453

ABSTRACT

<p><b>OBJECTIVE</b>To identify the mutation of the methylmalonic aciduria (cobalamin deficiency) CblC type, with homocystinuria (MMACHC) gene in a pedigree with methylmalonic aciduria.</p><p><b>METHODS</b>The MMACHC gene mutation was detected using polymerase chain reaction (PCR) and DNA sequencing. The MMACHC gene of 50 healthy people was also sequenced as control.</p><p><b>RESULTS</b>A new mutation of 146_154 del CCTTCCTGG was found in the patient and his father, and was absent in the controls.</p><p><b>CONCLUSION</b>A new mutation (146_154 del CCTTCCTGG) in the MMACHC gene was detected in a Chinese family with methylmalonic aciduria.</p>


Subject(s)
Animals , Child, Preschool , Female , Humans , Male , Pregnancy , Amino Acid Metabolism, Inborn Errors , Genetics , Metabolism , Amino Acid Sequence , Base Sequence , Carrier Proteins , Chemistry , Genetics , Case-Control Studies , DNA Mutational Analysis , Exons , Genetics , Fathers , Methylmalonic Acid , Metabolism , Molecular Sequence Data , Mutation , Pedigree , Polymerase Chain Reaction , Protein Structure, Secondary
7.
Chinese Journal of Biotechnology ; (12): 241-245, 2007.
Article in Chinese | WPRIM | ID: wpr-325386

ABSTRACT

Based on the sequence of BAC (Bacterial Artificial Chromosome) along with the Cre/lox P system, the gene-targeting vectors to multiple loci of the repetitive internal transcribed spacers between rDNA genes in Leghorn chicken were constructed. The key material of multiple loci gene targeting in vivo would be obtained. First, the plasmid of pYLSV-TDN with TK, HRDS2, and Neo genes was constructed. The TK-HRDS2-Neo DNA fragment obtained from the plasmid of pYLSV-TDN was digested by Not I/HindIII and inserted into the upstream of the lox P site of BAC plasmid for obtaining the selective vector of BAC-TDN. The expression vector of pYLVS-GID with EGFP, hIFN genes, and HRDS1 was then obtained. The plasmid of BAC-TDN-VS-GID was obtained by cotransformation of the selective vector of BAC-TDN and the expression vector of pYLVS-GID to E. coli NS3529 through the action of Cre/lox P system. The gene-targeting vector of BAC-TDN-GID to multiple loci of the ITS region in Leghorn chicken was obtained by cleaving the sequence of pYLVS with the homing endonuclease of I -Sce I and ligating with the linker of LS. The insertion and the insert direction of DNA fragments were identified by restriction digestion or PCR and sequencing in each clone. The significance of the technique ofgene-targeting vector to multiple loci are shown as follows. First, the targeting loci were increased to 100 - 300. Second, the problems of unstable expression of inserted genes were partially solved. Third, the need for safety against toxicity integration was resolved. Fourth, the forbidden zone of gene integrating on the repetitive DNA sequences was broken through.


Subject(s)
Animals , Humans , Attachment Sites, Microbiological , Genetics , Chromosomes, Artificial, Bacterial , Genetics , Cloning, Molecular , DNA , Genetics , Metabolism , DNA Restriction Enzymes , Metabolism , DNA, Ribosomal Spacer , Genetics , Escherichia coli , Genetics , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Integrases , Genetics , Interferon-gamma , Genetics , Polymerase Chain Reaction , Recombinant Fusion Proteins , Genetics , Transformation, Genetic
8.
Chinese Journal of Biotechnology ; (12): 125-130, 2006.
Article in Chinese | WPRIM | ID: wpr-237013

ABSTRACT

mPem, a homeobox gene, is expressed in a time and stage specific manner during murine ontogeny. Pem transcripts are abundant in 7- and 8-day mouse embryos, but decrease precipitously thereafter. On Day 9 they become abundant in placenta and yolk sac, persisting there until parturition. Although Pem transcripts are not detectable in most of adult tissues, they are present in reproductive system such as testis, epididymis and ovary. This indicates a important role for Pem during embryogenesis and reproductive development. To study the function of mPem protein, we used a GAL4 based yeast two-hybrid assay to screen a 7-day mouse embryo library with full-length of mPem. 3 proteins were found interacting with mPem protein. One of theses is Mdfic. We confirmed the interaction between mPem and Mdfic in yeast and in vitro. Mdfic, MyoD family inhibitor domain containing, encodes the myoD family inhibitor domain (I-mfa domain). The interaction between mPem and Mdfic suggested they maybe form the transcriptional regulator complex to regulate embryo differentiation.


Subject(s)
Animals , Female , Mice , Pregnancy , Embryo, Mammalian , Embryonic Development , Genes, Homeobox , Genetics , Homeodomain Proteins , Chemistry , Metabolism , Protein Binding , Transcription Factors , Chemistry , Metabolism , Two-Hybrid System Techniques
9.
Chinese Journal of Biotechnology ; (12): 996-1001, 2006.
Article in Chinese | WPRIM | ID: wpr-325436

ABSTRACT

VPAC2 is a co-receptor of pituitary adenylate cyclase activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) and mediates multiple bio-functions. In order to construct the CHO line expressing VPAC2 stably, pcDNA-VPAC2 was used to transfect CHO cells. The positive clones were selected by G418 and the clone VPAC2-CHO with high sensitivity to PACAP38 was picked out by its ability to promoting the concentration of cAMP. RT-PCR, Western blot and Immunofluorescenece assay were used to identify the express of VPACS. Binding competition with VPAC2 agonist and the bioactivity of mediating the ligand to promote the concentration of cAMP showed that VPAC2 was expressed effectively in VPAC2-CHO. The results of Scatchard analysis revealed that VAPC2-CHO expressed a receptor density of (1.1 +/- 0.2) pmol/mg protein, respectively, with Kd values of (0.55 +/- 0.10) nmol/L for PACAP38 used as a tracer. The construction of CHO cells expressing VPAC2 specially and functionally lays a foundation not only for the further research on the characters and functions of VPAC2 but also for the screening and characterization of novel agonists of antagonists for VPAC2.


Subject(s)
Animals , Cricetinae , Binding, Competitive , CHO Cells , Cell Membrane , Metabolism , Cricetulus , Cyclic AMP , Metabolism , Gene Expression , Genetic Vectors , Genetics , Iodine Radioisotopes , Chemistry , Pituitary Adenylate Cyclase-Activating Polypeptide , Chemistry , Metabolism , Pharmacology , Receptors, Vasoactive Intestinal Peptide, Type II , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Transfection
10.
China Biotechnology ; (12)2006.
Article in Chinese | WPRIM | ID: wpr-686458

ABSTRACT

Human cytomegalovirus (HCMV) is extremely species specific and does not replicate in experimental animal tissues.To overcome the problem and establish suitable animal models for studying antiviral strategies,the expression of HCMV UL49 gene was explored in mice.UL49-GFP gene was subcloned into the adenovirus shuttle plasmid pDC316,the products(pDC316-UL49-GFP)were co-transfected with helper plasmid pBHGloxE1,3Cre into HEK293 cell lines by liposome reagent,recombinant adenovirus(Ad-UL49-GFP) was generated and confirmed by PCR and Western blot.Ad-UL49-GFP was propagated in 293 cells and purified.The titer of viral stocks was determined by end-point dilution assay.The purified adenoviruses were delivered into mice via the tail vein injection.Fluorescence quantitative PCR and Western blot experiments were used to examine the tissue distribution and duration of UL49 gene expression.The results showed that the recombinant adenovirus were present in vivo.The expression level in tissues arranged in descending order was liver,spleen,kidney,heart and lung.3 days after injection,the liver,spleen,kidney,heart and lung expressed protein UL49 in high lever and then declined gradually.14 days after injection,UL49 protein expression was disappear in some organs except liver and spleen.In conclusion,transgene animal model carrying UL49 gene was successfully established.Therefore,the system may be suitable for selecting anti-HCMV drugs targeting UL49 gene.

11.
Microbiology ; (12)1992.
Article in Chinese | WPRIM | ID: wpr-684691

ABSTRACT

Human cytomegalovirus (HCMV) is a DNA virus and serious opportunistic pathogen for both newborn and immunocompromised individuals.To research technique for gene silence and antiviral agents, ribozyme M1GS-T6 was constructed from external guide sequences(EGSs)that consist of a sequence complementary to HCMV UL54 gene RNA and M1 RNA, the catalytic RNA subunit of RNase P from Escherichia coli. The results showed that M1GS can efficiently cleave the mRNA sequence encoding UL54 protein in vitro.

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