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Objective: To express DNA-binding protein (DBP) of human adenovirus (HAdV) type 7 using the prokaryotic expression system, and product anti-HAdV-7 DBP rabbit polyclonal antibody. Methods: The HAdV-7 DBP gene was synthesized and cloned into prokaryotic expressing vector pET30a, and the recombinant plasmid was transformed into E. coli BL21 (DE3) competent cell. The recombinant protein DBP was expressed by induced Isopropyl-beta-D-thiogalactopyranoside (IPTG) and purified with Ni-NTA affinity column. The titer of anti-DBP polyclonal antibody produced in immunized rabbit was measured by indirect ELISA, and the specificity of the antibody was identified by Western blotting and indirect immunofluorescence assay (IFA). In addition, purified rDBP was used as coating antigen for indirect ELISA assay to detect specific IgM and IgG antibodies against DBP in the serum of children infected with HAdV. Results: The HAdV-7 DBP plasmid was constructed successfully. The purified recombinant DBP was more than 95% after purification. The titer of polyclonal antibody was 1∶1 024 000. The polyclonal antibody showed high specificity in vitro using Western blotting and IFA. The positive rate of specific anti-DBP IgM and IgG antibody in acute-phase serum samples collected from children infected with HAdV were 50.0% (19/38) and 63.2% (24/38), respectively, using indirect ELISA. Conclusion: In summary, the HAdV-7 rDBP is expressed using prokaryotic expression system, and the recombinant HAdV-7 DBP protein and the anti-DBP rabbit polyclonal antibody with high titer are prepared.
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Animals , Rabbits , Adenoviruses, Human/genetics , Antibody Specificity , Blotting, Western , DNA-Binding Proteins/metabolism , Enzyme-Linked Immunosorbent Assay , Escherichia coli/genetics , Immunoglobulin GABSTRACT
Objective To observe the effect of urantide on the expression of osteopontin (OPN) and α-smooth muscle actin (a-SMA) in the heart tissue of atherosclerosis (AS) rats, and to explore its mechanism of prevention and treatment of myocardial fibrosis injury in rats. Methods Totally 120 3-week-old healthy male Wistar rats in SPF grade were randomly divided into six groups; control group, model group, simvastatin group, urantide (3 days, 7 days, 14 days). HE and Masson trichrome staining were used to observe the morphology of rat heart and the expression of collagen fibers. Immunohistochemistry and Western blotting were used to detect the expression of OPN and α-SMA protein. Results In AS model group, cardiomyocyte hypertrophy or atrophy, a large number of inflammatory cell infiltration and a small amount of foam cells were observed in the heart tissue of rats. The increase of collagen fibers and the expression of OPN and α-SMA protein in cardiac tissue were significantly higher than those in the control group. Compared with the AS model group, after urantide treatment, cardiac injury was significantly improved, and the expression of collagen fiber, OPN and α-SMA protein was decreased. Conclusion Urantide can inhibit the expression of OPN and α-SMA protein in the heart tissue of AS rats to alleviate myocardial fibrosis and play a protective role in the heart tissue of AS rats.
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Bronchiolitis is an acute lower respiratory tract infection in early childhood. Respiratory syncytial virus(RSV)is the most common etiology of bronchiolitis. Infections with Mycoplasma pneumoniae,Chlamydia pneumonia and Chlamydia trachomatis can also cause bronchiolitis. Bronchiolitis mainly occurs in infants under 2 years of age,with a peak age of 2 to 6 months. Bronchiolitis is the most common reason for hospitalization in infants,and infants under 6 months of age have the highest hospitalization rate. Since bronchiolitis is mainly caused by RSV infection,its seasonal characteristics are similar to those of local RSV infection. Bronchiolitis is a disease with a high incidence but a low mortality rate. Being less than 3 months,prematurity,congenital heart disease and so on are risk factors of severe bronchiolitis.
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Viruses are the main causes of respiratory tract infection in children. Early,rapid and accurate diagnosis of pathogen in respiratory tract infection can avoid antibiotic abuse.There are advantages and disadvantages of different laboratory diagnostic methods for respiratory virus infections. Thus,appropriate choice of diagnostic methods and correct interpretation of the test results can help clinicians to make accurate pathogen diagnosis.
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Objective To analyze the clinical characteristics of community-acquired influenza virus pneumonia in hospitalized children and improve the clinicians' understanding level of this disease.Methods Data of 70 cases with community-acquired influenza virus pneumonia admitted to the Respiratory Department and Infectious Disease,Beijing Children's Hospital,Capital Medical University,from November 2009 to April 2018 were collected and the clinical characteristics were analyzed.Results Of the 70 cases,61 cases(89.7％) were discharged after improvement.The median age was 3.5 years old,and 50 cases(71.4％) were 0 to 5 years old.There were 29 cases with severe influenza pneumonia,41 cases with mild influenza pneumonia,3 cases died,and 19 cases (27.1％) had underlying diseases.Sixty-four cases (91.4％) were hospitalized in winter and spring.The first symptoms were mainly fever in 64 cases (91.4％) and cough in 65 cases (92.9％),and temperatures were mostly from 39.1 ℃ to 41.0 ℃.Lung auscultation was dominated by moist rales (30 cases,58.8％) and wheezing (8 cases,15.7％).There were many complications of influenza virus pneumonia,including 19 cases with myocardial injury,11 cases with liver function injury,4 cases with toxic encephalopathy,3 cases with electrolyte disturbance,2 cases with multiple organ failure,2 cases with hemophagocytic syndrome,and 1 case with septic shock.Chest radiographic results reveal bilateral inflammation in 40 children (57.1％),prodominatly in lower lobe lesions (39 cases).The common changes were patchy shadow,interstitial parenchymal lesion,ground glass shadow,and pleural effusion.Forty-seven children (67.1％) were infected by influenza A,and 23 children(32.9％) were co-infected.The percentage of severe cases with underlying diseases (68.4％) was significantly higher than that in children without chronic diseases (31.4％),the difference was statistically significant (x2 =7.830,P =0.005).The increase rate of C reaction protein (CRP) in severe cases (54.3％) was significantly higher than that in mild cases (28.6％),the difference was statistically significant (x2 =4.769,P =0.029).Conclusions Community-acquired influenza virus pneumonia in children mainly occurs in winter and spring.It is more common seen in children under 5 years of age.The main clinical manifestations of community-acquired influenza virus pneumonia are high fever and cough,extrapulmonary complications are more common.Most children have moist rales and showed bilateral inflammation and lower lobe lesions in chest radiography.Children with underlying diseases are more likely to develop severe influenza virus pneumonia.Elevated CRP is associated with severe influenza virus pneumonia.Most patients have a good prognosis,but there are still cases of death.
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The study aims at developing a convenient and specific method for the identification of Fel Serpentis DNA. The methods of Fel Serpentis genomic DNA purification were tested and optimized, four pairs of specific primers for the amplification of COⅠ, Cyt b and 16S were designed. Then the best pair of primers were selected according to the specificity and efficiency. The DNA fragment about 400 bp was amplified from 20 kinds of Fel Serpentis, whereas no DNA fragment was amplified from other animal samples under the same condition. This method is specific,accurate and reproducible, which provides a useful tool for the quality control of Fel Serpentis.
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The study aims at investigating the ecological environment Adiantum reniforme var. sinense of in Three Gorges Reservoir region, and providing a reference basis for the protection of resources and artificial cultivation of A. reniforme var. sinense. By using the method of investigation, field survey and experimental analysis, the vegetation, natural geographical environment, climate, soil nutrients of A. reniforme var. sinense were studied and analyzed. The survey found that A. reniforme var. sinense distribution area reduced fast in Three Gorges region, a lot of distribution has diminished and vanished due to excessive digging, currently only in 3 towns of Wanzhou there exist 4 wild distribution areas. The growth of A. reniforme var. sinense needs an environment with low altitude, steep slope and thin soil, northeast slope, canopy height and warm and humid climate characteristics, and the soil in distribution has the characteristics of high organic matter, available nitrogen, available potassium, and low available phosphorus content.
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Objective To observe the effects of Wendan decoction (WD) on depression-like behavior in a rat model of Parkinson's disease (PD) and explore the related mechanism.Methods Rodent model of PD was established by unilaterally lesioning medial forebrain bundle with 6-hydrodopamine.After intragastric administration with WD,the rats's behavior changes were detected by the open field test,sucrose preference test and forced swimming test;the contents of monoamine neurotransmitters in the rat brain were assessed by high-performance liquid chromatography with electrochemical detection.Results Compared with those of sham-operated rats,the horizontal and vertical activities of the PD model rats decreased significantly,and sucrose consumption decreased significantly,but immobility time during forced swimming was significantly prolonged.The contents of dopamine (DA),5-hydroxytryptamine (5-HT) and noradrenaline (NA) in the medial prefrontal cortex (mPFC) and DA in the striatum decreased significantly.After administration of WD for 2 weeks,the immobility time of the PD model rats was significantly decreased,sucrose consumption increased significantly;DA,5-HT and NA levels in the mPFC increased significantly.Conclusion WD improves the depression-like behavior in PD model rats,and the mechanisms may involve the regulation of monoamine neurotransmitters in mPFC.
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Objective To study correlation between the deformation and displacement trend of the lumbosacral vertebra (L1-S1) for two typical scoliosis spine under vertical load, so as to provide the mechanical basis of treatment and prevention of scoliosis in clinic. Methods The X-ray computed tomography (CT) imaging of two typical scoliosis spine (Lenke-4AN type and Lenke- 5CN type) were converted into 3D models，and their finite element models were then established and verified. The internal stress distribution and displacement variation of the models were calculated by the finite element software; the correlation between the lumbosacral vertebral structure and displacement of the spine was analyzed. Results Under the same boundary conditions and load cases, the stress and displacement for two kinds of lumbosacral vertebral models showed different trends. Due to its left-leaning and forward convex bending deformation as well as relatively large lordosis angle(60°) and smaller left-leaning angle (17.37°), the Lenke-4AN type lumbosacral spine produced slightly small forward convex displacement(8.18 mm) and relatively large left-leaning displacement (0.97 mm). The Lenke- 5CN type lumbosacral spine showed left-leaning and forward convex bending deformation as well, with relatively large lordosis angle (59°) and left-leaning angle (26.97°), so it produced more severe left-leaning (20.65 mm) and forward deformation (9.22 mm). Conclusions The deformation trend of lumbosacral vertebra is closely related to its structural characteristics, and different scoliosis lumbosacral vertebral structures will cause corresponding deformation trend. The research findings are important for the prevention and treatment of scoliosis.
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Objective To launch systematic research on long-term asymptomatic hyperuricemia (HUA) from hemorheological viewpoint, so as to provide references for clinical treatment of asymptomatic HUA. Methods Twenty rats were randomly and evenly divided into normal control group and model group. The rats were intraperitoneally injected with 250 mg/(kg•d) oxonate for 8 weeks to induce the model of asymptomatic HUA. The blood samples were obtained to measure the serum uric acid, hemorheological parameters, oxidative and anti-oxidative indices. Results The aggregation index, haemolysis rate, serum xanthine oxidase (XOD), plasma fibrinogen and blood viscosity significantly increased, while the orientation index, electrophoresis rate, serum superoxide dismutase (SOD), activated partial thromboplastin time (APTT) and prothrombin time (PT) significantly induced. Conclusions The asymptomatic HUA can lead to more serious oxidative stress, deteriorate the hemorheological parameters of red blood cells in rats, and induce higher blood viscosity and coagulation status. The research findings indicate that asymptomatic HUA should be correctly understood and timely intervened in clinical diagnosis.
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Objective To study the correlation between the deformation and displacement trend of the lumbosacral vertebra (L1-S1) for two typical scoliosis spines under vertical loads,so as to provide the mechanical basis of treatment and prevention of scoliosis in clinic.Methods The X-ray computed tomography (CT) images of two typical scoliosis spines (Lenke-4AN type and Lenke-5CN type) were converted into 3D models,and their finite element models were then established and verified.The internal stress distribution and displacement variation of the models were calculated by the finite element software;the correlation between the lumbosacral vertebral structure and displacement of the spine was analyzed.Results Under the same boundary conditions and load cases,the stress and displacement for two kinds of lumbosacral vertebral models showed different trends.Due to its leftleaning and forward convex bending deformation as well as the relatively large lordosis angle (60°) and smaller left-leaning angle (17.37°),the Lenke-4AN type lumbosacral spine produced slightly small forward convex displacement (8.18 mm) and relatively large left-leaning displacement (0.97 mm).The Lenke-5CN type lumbosacral spine showed left-leaning and forward convex bending deformation as well,with relatively large lordosis angle(59°) and left-leaning angle (26.97°),so it produced more severe left-leaning displacement (20.65 mm) andforward convex displacement (9.22 mm).Conclusions The deformation trend of lumbosacral vertebra is closelyrelated to its structural characteristics,and different scoliosis lumbosacral vertebral structures will cause the corre-sponding deformation trend.The research findings are important for the prevention and treatment of scoliosis.
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Objective To launch systematic research on long-term asymptomatic hyperuricemia (HUA) from hemorheological viewpoint,so as to provide references for clinical treatment of asymptomatic HUA.Methods Twenty rats were randomly and evenly divided into normal control group and model group.The rats were intraperitoneally injected with 250 mg/(kg · d) oxonate for 8 weeks to induce the model of asymptomatic HUA.The blood samples were obtained to measure the serum uric acid,hemorheological parameters,oxidative and anti-oxidative indices.Results The aggregation index,haemolysis rate,serum xanthine oxidase (XOD),plasma fibrinogen and blood viscosity significantly increased,while the orientation index,electrophoresis rate,serum superoxide dismutase (SOD),activated partial thromboplastin time (APTT) and prothrombin time (PT) significantly induced.Conclusions The asymptomatic HUA can lead to more serious oxidative stress,deteriorate the hemorheological parameters of red blood cells in rats,and induce higher blood viscosity and coagulation status.The research findings indicate that asymptomatic HUA should be correctly understood and timely intervened in clinical diagnosis.
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Objective To study the correlation between the deformation and displacement trend of the lumbosacral vertebra (L1-S1) for two typical scoliosis spines under vertical loads,so as to provide the mechanical basis of treatment and prevention of scoliosis in clinic.Methods The X-ray computed tomography (CT) images of two typical scoliosis spines (Lenke-4AN type and Lenke-5CN type) were converted into 3D models,and their finite element models were then established and verified.The internal stress distribution and displacement variation of the models were calculated by the finite element software;the correlation between the lumbosacral vertebral structure and displacement of the spine was analyzed.Results Under the same boundary conditions and load cases,the stress and displacement for two kinds of lumbosacral vertebral models showed different trends.Due to its leftleaning and forward convex bending deformation as well as the relatively large lordosis angle (60°) and smaller left-leaning angle (17.37°),the Lenke-4AN type lumbosacral spine produced slightly small forward convex displacement (8.18 mm) and relatively large left-leaning displacement (0.97 mm).The Lenke-5CN type lumbosacral spine showed left-leaning and forward convex bending deformation as well,with relatively large lordosis angle(59°) and left-leaning angle (26.97°),so it produced more severe left-leaning displacement (20.65 mm) andforward convex displacement (9.22 mm).Conclusions The deformation trend of lumbosacral vertebra is closelyrelated to its structural characteristics,and different scoliosis lumbosacral vertebral structures will cause the corre-sponding deformation trend.The research findings are important for the prevention and treatment of scoliosis.
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Objective To launch systematic research on long-term asymptomatic hyperuricemia (HUA) from hemorheological viewpoint,so as to provide references for clinical treatment of asymptomatic HUA.Methods Twenty rats were randomly and evenly divided into normal control group and model group.The rats were intraperitoneally injected with 250 mg/(kg · d) oxonate for 8 weeks to induce the model of asymptomatic HUA.The blood samples were obtained to measure the serum uric acid,hemorheological parameters,oxidative and anti-oxidative indices.Results The aggregation index,haemolysis rate,serum xanthine oxidase (XOD),plasma fibrinogen and blood viscosity significantly increased,while the orientation index,electrophoresis rate,serum superoxide dismutase (SOD),activated partial thromboplastin time (APTT) and prothrombin time (PT) significantly induced.Conclusions The asymptomatic HUA can lead to more serious oxidative stress,deteriorate the hemorheological parameters of red blood cells in rats,and induce higher blood viscosity and coagulation status.The research findings indicate that asymptomatic HUA should be correctly understood and timely intervened in clinical diagnosis.
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This study aims to investigate the genetic characteristics of BZLF1 gene and its promoter Zp of the epidemic strains in children with primary Epstein-Barr virus (EBV)-associated diseases. Total DNA was extracted from the peripheral blood of 134 children with EBV-associated infectious mononucleosis (EBV-IM) and 32 children with EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH) who were admitted to Beijing Children's Hospital from 2006 to 2011. The EBNA3C, BZLF1, and Zp genes were amplified by PCR assay. Typing of EBV was performed according to the size of the amplification product of EBNA3C gene; the amplification products of BZLF1 and Zp genes were subjected to direct sequencing, and sequence analysis was performed using BioEdit 7. 0. 9. The results were as follows: (1) EBV-1 was present in 140 samples (97.2%, 140/144) and EBV-II in 4 samples (2.8%, 4/144). (2) Three BZLF1 genotypes and their 12 subtypes (including 6 newly found subtypes) were detected in this study; there were no significant differences in the frequencies of BZLF1-A and BZLF1-B between the children with EBV-IM and EBV-HLH (P = 0.083); BZLF1-A1 was the dominant genotype in children with EBV-associated diseases; t BZLF1-A mostly had three 29-bp repeats in the first intron of BZLF1 gene, and BZLF1-B mostly had 30-bp repeats (P = 0.000), with the number of repeats varying from 1 to 13. (3) Four Zp genotypes were detected in this study, including Zp-P, Zp-V3, Zp-V4, and Zp-V1; there were no significant differences in the frequencies of these Zp genotypes between children with EBV-IM and EBV-HLH (P = 0.272, 0.252, 1.0, and 1.0, respectively). (4) The linkage analysis of BZLF1 gene and its promoter Zp showed that BZLF1-A1 was highly associated with Zp-V3 (P = 0.000), while BZLF1-B4 with Zp-P (P = 0.000); EBV-I + BZLF1 A1 was highly associated with Zp-V3 (P = 0.000), while EBV-I+BZLF1-B4 with Zp-P (P = 0.000). The conclusions are as follows: (1) BZLF1-A1 is the dominant genotype in children with EBV-associated diseases; there are mostly 29-bp repeats in the first intron of BZLF1 gene for BZLF1-A genotype and 30-bp repeats for BZLF1-B genotype. (2) Zp-P and Zp-V3 are dominant Zp genotypes of EBV in children, which shared similar detection rates. (3) BZLF1-A1 is highly associated with Zp-V3, while BZLF1-B4 with Zp-P; EBV-I+BZLF1-A1 is highly associated with Zp-V3, while EBV-I+BZLF1-B4 with Zp-P.
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Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , China , Epidemiology , Epstein-Barr Virus Infections , Epidemiology , Virology , Genotype , Herpesvirus 4, Human , Genetics , Physiology , Introns , Genetics , Promoter Regions, Genetic , Genetics , Repetitive Sequences, Nucleic Acid , Genetics , Trans-Activators , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To measure the expression of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (PKB) in liver tissue among low-birth-weight newborn rats treated with L-arginine (L-Arg) in early life, and to investigate the effect of L-Arg on insulin resistance.</p><p><b>METHODS</b>Eighteen pregnant rats were randomly divided into three groups: control, model and intervention (n=6 each). The control group was fed with normal protein feed (protein content=21%) during pregnancy to establish a normal-birth-weight newborn rat model, and the model and intervention groups were fed with low-protein feed (protein content=10%) during pregnancy to establish a low-birth-weight newborn rat model. Newborn rats from the three pregnant rat groups were also assigned to control, model and intervention groups. During 21 days of lactation, maternal rats in the control and model groups were fed with normal protein feed and normal drinking water, while maternal rats in the intervention group were fed with normal protein feed and drinking water rich in L-Arg (200 mg/kg·d). After ablactation, the three groups of newborn rats were fed with normal protein feed and normal drinking water. Liver tissue samples were collected from these newborn rats at 1, 3 and 8 weeks after birth. Protein expression of PI3K and PKB in liver tissue was measured by Western blot.</p><p><b>RESULTS</b>At 1 week after birth, the newborn rats in the intervention group had significantly higher protein expression of PI3K than in the model group (P=0.045), but there was no significant difference when compared with the control group (P=0.503). At 8 weeks after birth, the newborn rats in the intervention group had significantly higher protein expression of PKB than the model group (P=0.039), but there was no significant difference when compared with the control group (P>0.05).</p><p><b>CONCLUSIONS</b>A supplement of L-Arg in early life can boost protein synthesis, increase protein expression of PI3K and PKB in liver tissue, promote insulin signaling and reduce insulin resistance.</p>
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Animals , Female , Male , Pregnancy , Rats , Animals, Newborn , Arginine , Pharmacology , Birth Weight , Liver , Metabolism , Phosphatidylinositol 3-Kinases , Genetics , Phosphorylation , Proto-Oncogene Proteins c-akt , Genetics , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To introduce the experience in the treatment of lower eyelid pouches orbital rim.</p><p><b>METHODS</b>An incision was made along the margin of lower eyelid and dissection was performed under the orbicularis muscle to expose the orbital septum and periosteum of lower orbital rim. The fat released from orbital septum was transposed just below the lower orbital rim and fixed on the periosteum. If lacrimal groove deformity was not corrected completely, the musculocutaneous flap, which may be excised beside the incision, was kept to correct the deformities further with only the muscle portion.</p><p><b>RESULTS</b>72 cases with lower eyelid pouches complicated with lacrimal groove deformities were treated with transposition of orbital fat and orbicularis muscular flaps. Satisfactory results were achieved in all the patients after a follow-up period of 3-6 months.</p><p><b>CONCLUSION</b>It is an effective and feasible technique to correct lacrimal groove deformities with transposition of orbital fat and orbicularis muscular flaps.</p>
Subject(s)
Aged , Humans , Adipose Tissue , Transplantation , Blepharoplasty , Methods , Eyelids , General Surgery , Orbit , Periosteum , General SurgeryABSTRACT
<p><b>OBJECTIVE</b>Acute respiratory tract infections (ARI) are the leading cause of pediatric morbidity and mortality worldwide, particularly in developing countries. Viruses are the main pathogens of ARI in children. The purpose of the present study was to determine the epidemiologic features of respiratory viruses, including novel viruses, in outpatient and hospitalized children with ARI.</p><p><b>METHOD</b>From March 2010 to February 2012, 2066 children with ARI, including 1050 outpatients and 1016 inpatients, were involved in this study. One nasopharyngeal aspirate or throat swab specimen was collected from each patient. Reverse transcription (RT) PCRs were performed to detect common respiratory tract viruses including respiratory syncytial virus (RSV), human rhinovirus (HRV), influenza virus (IFV), parainfluenza virus (PIV) type 1-4, adenovirus (ADV), enterovirus (EV), human coronavirus (HCOV), human metapneumonia virus (HMPV) and human bocavirus (HBOV).</p><p><b>RESULT</b>At least one viral pathogen was identified in each of 1274 out of 2066 patients and the overall positive rate was 61.7%. The positive rate in inpatient (69.7%) was higher than that in outpatient (53.9%). The frequencies of detection of various viruses among in- and outpatients were different. RSV was the most prevalent virus detected among hospitalized children, followed by HRV and PIV, whereas IFV was the most frequently identified virus in the outpatient group, followed by ADV and PIV. Simultaneous detection of two or more viruses was found in 377 cases. Coinfection was more frequent in inpatients than in outpatients (30.1% vs. 6.8%, P < 0.001).</p><p><b>CONCLUSION</b>Respiratory viruses play an important role in children with ARI, especially in young children. RSV was the most prevalent virus detected among hospitalized children, whereas IFV was the most frequently identified virus in the outpatient group. Viral coinfections are frequently identified, particularly in hospitalized patients. Further studies are required to better understand the impact of coinfections in children with ARI.</p>
Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Male , Acute Disease , Age Distribution , Child, Hospitalized , China , Epidemiology , Coinfection , Epidemiology , Virology , DNA Viruses , Nasopharynx , Virology , Outpatients , Parainfluenza Virus 1, Human , Parvoviridae Infections , Epidemiology , Respiratory Syncytial Virus Infections , Epidemiology , Respiratory Syncytial Virus, Human , Respiratory Tract Infections , Epidemiology , Virology , Reverse Transcriptase Polymerase Chain Reaction , Rhinovirus , SeasonsABSTRACT
Objective To investigate the genetic polymorphism of 13 cytokines genes in Han individual children from Beijing area.Methods The polymerase chain reaction-sequence specific primers (PCR-SSP) method was used to determine cytokine gene polymorphism including interleukin (IL)-1α (-889C/T),IL-1β (-511C/T,+3942T/C),IL-1 R(pst1 1970C/T),IL-1RA(mspa1 11100T/C),IL-4Ra(+ 1902G/A),IL-12 (-1188C/A),interferon (IFN)-γ(+ 874A/T),transforming growth factor (TGF)-β1 (codon10C/T,codon25 G/C),tumor necrosis factor)(TNF)-α (-308G/A,-238G/A),IL-2 (-330T/G,+ 166G/T),IL-4 (-1098 T/G,-590 T/C,-33 T/C),IL-6(-174G/C,nt565G/A),IL-10 (-1082G/A,-819C/T,-592A/C) 13 cytokines genes in 210 unrelated healthy Han individuals from Beijing area.Results (1) Cytokine genes with frequencies more than or equal to 95 percent included IL-1β +3962 C,TGF-β Codon 25 G,IL-6-174 G,IL-6 nt565 G,TNF-α-308 G and TNF-α-238 G alleles;while cytokine genes with frequencies less than or equal to 5 persent included IL-1β + 3962 T,TGF-β Codon 25 C,IL-6-174C,IL-6 nt 565 A,TNF-α-308 A and TNF-α-238 A alleles.(2) The frequency of the high-producer-type GCC haplotype in IL-10 gene was 8.27％,while the low-producer-type ATA was 59.11％.The frequency of the highproducer-type TG haplotype in the TGF-β gene was 49.77％,while the low-producer-type CC was 0.The frequencies of the high-producer-types AG and AA haplotypes in the TNF-α gene were 3.34％,while those of the low-producer-types GG and GA were 96.67％.The frequency of the high-producer-types T/G genotype in the IL-2-330 gene was 11.90％,while those of the low-producer-types T/T and T/G were 88.10％.The frequencies of the high-producer-types G/G and G/C genotypes in the IL-6-174 gene were 99.52％,while that of the low-producer-types C/C was 0.48％.The frequency of the high-producer-types T/T genotype in the INF-γ+ 874 gene was 1.90％,and the frequency of the medium-producer-types T/A in the INF-γ+ 874 gene was 22.86％,while of the low-producer-types A/A was 75.24％.(3) New genotypes were found:the frequency of IL-2(-330,+ 166) GG/GT/TT genotype was 1.90％,and the frequency of IL-2(-330,+ 166) TG/GG/GT/TT genotype was 0.48％,The frequency of IL-6(-174,nt565) CG/CG genotype was 0.48％,the frequency of IL-6 (-174,nt565) GA/GG genotype was 1.90％,The frequency of IL-6 (-174,nt565) GG/GA/CA genotype was 0.48％,The frequency of IL-10(-1082,-819,-592) GCC/ATC/ATA genotype was 0.95％,The frequency of IL-10(-1082,-819,-592) ACC/ACA/ATC/ATA genotype was 35.71％,The frequency of IL-10 (-1082,-819,-592) GCC/ACC/ACA/ATC/ATA genotype was 0.48％.Conclusion The genetic polymorphism of cytokine genes in Han individuals from Beijing area is different from other areas of China with higher frequencies of IL-10 (-1082,-819,-592) ACC/ACA/ATC/ATA genotype.
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<p><b>OBJECTIVE</b>To analyze the characteristic of nuclear antigen 1 gene and latent membrane protein 1 gene of Epstein-Barr virus in primary EBV infection in children in Beijing area in 2005-2012.</p><p><b>METHODS</b>Polymerase chain reaction (PCR) was used to amplify the EBNA-3C, EBNA1 and LMP1 genes. The amplified products were sequenced directly and the sequences were analyzed by BioEdit 7. 0. 9 and MEGA 4. 0. 2.</p><p><b>RESULTS</b>Type A EBV was detected in 98% samples. Nucleotide sequence analysis of the carboxy-terminal region of EBNA1 showed that Vvvl was deteted in 98% samples. DNA sequence analysis of LMP1 C-terminus indicated that China 1 was 90% in this study. There were no significant differences in the frequency of Vvv1 and China 1 between the IM and HLH samples (P = 1.00). Linkage analysis of EBV types, EBNA1 and LMP1 variants indicated that 90% of EBV type A was associated with EBNA1-Vvv1 variant and LMP1-China 1 variant in 40 cases. Full length of LMP1 gene was successfully amplified in 35 cases. Four Chinese groups (CG1-4) were identified. The percentage of CG1-CG4 were 85%, 6%, 6% and 3%, respectively.</p><p><b>CONCLUSION</b>EBV type A is predominant in primary EBV infection in children in Beijing Area. EBNA1-Vvv1 and LMP1-China 1 variants were predominant genotypes in this area. There is a high linkage between EBNA1-Vvv1 variant and LMP1-China 1 variant. Four Chinese groups (CG1-4) were identified according to the full length of LMP1 gene and CG1 was the most prevalent.</p>