Objective:To express Gag gp120 fusion protein in the recombinant fowlpox virus.Methods:The Gag gp120 gene of HIV 1 was inserted downstream of the combined promoter in pUTAL,an expressing plasmid was constructed.By transfecting the plasmid into CEF cells pre infected with FPV 282E 4 strain,a recombinant fowlpox virus vUTALG was obtained by selecting in the presence of BUdR and subsequent blue plaque screening.Results:Western blot showed that the recombinant virus expressed the Gag gp120 fusion protein in the infected CEF cells.The virus like particles formed by Gag protein were observed under electron microscope.Mice were immunized with the recombinant virus.The results showed the recombinant virus could induce HIV 1 specific antibody response.Conclusion:The recombinant fowlpox virus could express the Gag gp120 fusion protein.

Objective To construct multiple myeloma mucin MUC1-2VNTR gene eukaryotic expressing vector,which provided the basic material for further study of multiple myeloma DNA vaccine.Methods MUC1-2VNTR coding gene as target gene,and a KOZAK sequence was inserted before it.Hind Ⅲ and Xba Ⅰ restriction enzyme site were inserted on both ends.Then the whole sequence was synthesized and cloned into pcDNA3.1/myc-his B vector,and the recombinant vector was identified by restriction enzyme digestion and DNA sequencing.Results Synthesized MUC1-2VNTR gene was 140 bp.Restriction enzyme digestion and DNA sequencing confirmed pcDNA3.1/MUC1-2VNTR/myc-his B including the whole exact translation frame region and MUC1-2VNTR gene.Condnsion The pcDNA3.1/MUC1-2VNTR/myc-his B has been successfully constructed,which provides the basic material for further studies of MUC1 mucin function and multiple myloma DNA vaccine.

Objective To analyze karyotype,genetic characteristics and clinical features of Turner syn-drome.Methods Cytogenetic analysis and genetic counseling were performed for patients with the diagnosis of Turner syndrome.Results Analyzed karyotype of forty-nine patients with Turner syndrome,there are five major categories such as simple type,numerical abnormality and chimeras,structural abnormality and chime-ras,both numerical abnormality and structural abnormality chimeras,contained the Y chromosome.Abnormal karyotypes were presented at eighteen species.Simple type was the most prevalent type of patients with Turn-er syndrome(43%).Conclusion Different karyotype exhibit some different phenotype.The clinical manifesta-tion of chimeras might depend on the proportion of abnormal karyotype,Case contained the Y chromosome would be hermaphrodismy mostly.Timely diagnosis of the disease would have positive significance to preven-tion and therapy.Hormonal therapy could improve patient's height and gonad development,the case contained the Y chromosome should pay attention to prevent gonadoblastoma.

Based on the pathophysiology of the brain, advance in angiogenesis induced by stroke, and evidences of Chinese-medicine-mediated angiogenesis, the possibility to study the stroke-treating mechanism of Chinese medicine in angiogenesis was discussed. And regarding our previous work on angiogenesis modulated by qi-tonifying and stasis-eliminating therapy following intracerebral hemorrhage, we proposed some questions, which should be taken into account in the further work.
Brain ; blood supply ; pathology ; Humans ; Medicine, Chinese Traditional ; Microvessels ; pathology ; Neovascularization, Physiologic ; Stroke ; therapy ; Wound Healing

BACKGROUNDCC chemokine receptor 3 (CCR3), expressed on some inflammatory cells, is a member of the chemokine receptor family. Its ligand is eotaxin/CCL11. In this research, we studied the expression and function of CCR3 induced by interleukin-2 (IL-2) and interleukin-4 (IL-4) on human germinal centre (GC) B cells.

METHODSCells isolated from human tonsils were stimulated with IL-2 or/and IL-4 followed by bonding with eotaxin/CCL11. Flow cytometry was used to detect expression of CCR3 on GC B cells and apoptosis of GC B cells. Real time quantitative reverse transcription polymerase chain reaction and Northern blot assays were used to analyse the CCR3 mRNA expressed in the GC B cells. Chemotaxis and adhesion assays were used to determine the effect of eotaxin/CCL11 ligand bonded to CCR3 on GC B cells.

RESULTSThere was no CCR3 expression on human freshly isolated GC B cells. The combination IL-2 and IL-4 could upregulate CCR3 mRNA and protein expression on GC B cells. Eotaxin could not induce GC B cell chemotaxis and adhesion but triggered apoptosis of GC B cells.

CONCLUSIONIL-2 and IL-4 together induced expression of CCR3 on GC B cells, and the receptor acted as a death receptor.

Apoptosis ; B-Lymphocytes ; metabolism ; pathology ; Cell Adhesion ; Chemotaxis, Leukocyte ; Germinal Center ; metabolism ; pathology ; Humans ; Interleukin-2 ; pharmacology ; Interleukin-4 ; pharmacology ; RNA, Messenger ; analysis ; Receptors, CCR3 ; Receptors, Chemokine ; genetics

Adolescent ; Adult ; Aged ; Diagnostic Errors ; Female ; Fractures, Bone ; diagnosis ; Humans ; Male ; Middle Aged ; Multiple Trauma ; pathology ; therapy ; Physician's Role ; Young Adult

OBJECTIVETo investigate the proportion of incidence of children with non-specific chronic cough in Chongqing and analyze the characteristics of etiology during the follow-up.

METHODDiagnostic criteria were defined for children with non-specific chronic cough according to the Guidelines of diagnosis and therapy for children with chronic cough that were formulated by the Subspecialty Group, Society of Pediatrics, Chinese Medical Association and Chinese Journal of Pediatrics in 2008. Totally 266 patients in whom cough was the main or the only symptom,lasting > 4 weeks, presenting to Asthma Center of Children's Hospital, Chongqing Medical University between June 2008 and April 2009 were recruited into this study. Based on the Guidelines, diagnosis was made after taking history, physical examination and assistant examination. After etiological treatment, the patients were followed up during the second week, the fourth week and the twelfth week. Etiological diagnosis was confirmed if cough was resolved after specific therapy. If cough was not resolved,the diagnosis was rechecked and a new therapy was applied.

RESULTTotally 125 (47.0%) patients received final diagnoses of cough variant asthma (CVA), 58 (21.8%) was CVA and upper airway cough syndrome (UACS), 44 (16.5%) was diagnosed postinfection cough, 35 (13.2%) of UACS. In different age groups, the proportion of incidence of etiological agents is statistically distinct. In the ≤ 3 years old group, 35 patients (70.0%) were diagnosed CVA, 10 (20.0%) was postinfection cough; in 3 - 6 years group, 71 patients (50.7%) had CVA; the incidence of UACS was significantly higher in ≥ 6 years group.

CONCLUSIONIt is concluded that CVA, CVA and UACS, post infection cough, and simple UACS were identified as the three top reasons for children with chronic cough in Chongqing. Children with chronic cough of different age groups had different etiology of cough. The characteristic of each etiology need further study.

Adolescent ; Asthma ; epidemiology ; Child ; Child, Preschool ; China ; epidemiology ; Chronic Disease ; Cough ; epidemiology ; etiology ; microbiology ; Follow-Up Studies ; Humans ; Incidence ; Infant ; Infection ; epidemiology

OBJECTIVETo probe into the mechanism of electroacupuncture intervention for lipid metabolism of metabolic syndrome patients.

METHODSEighty cases of metabolic syndrome were randomly divided into an electroacupuncture combined with western medicine group (observation group) and a simple western medicine group (control group), 40 cases in each group. The observation group was treated with electroacupuncture at Back shu points, Zusanli (ST 36),Zhongwan (CV 12),Sanyinjiao (SP 6) etc. as main combined with oral administration of Simvastatin, Glipizide XL, and Felodipine sustained-release tablets for lipid-lowering, glucose-lowering and antihypertensive treatment; the control group was treated with oral administration of western medicine only (the medicine was the same with observation group). The Body Mass Index (BMI) and the blood lipid of the patients were detected respectively before and after treatment.

RESULTSThe BMI, Three Acids Glyceride (TG), Total Cholesterol(TC), Low Density Lipoprotein Cholesterol (LDL-C) and High Density Lipoprotein Cholesterol (HDL-C) were compared respectively before and after treatment, there were significant differences between them in observation group (all P < 0.01); while in control group, there were significant differences of TG,TC,LDL-C and HDL-C before and after treatment (all P < 0.01), and with no significant difference in BMI before and after treatment. There were significant differences of BMI,TG,TC,LDL-C and HDL-C between two groups after treatment (P < 0.01, P < 0.05).

CONCLUSIONThe electroacupuncture has an obvious effect to reduce body mass, and acupuncture combined with medication has a better effect of improving the lipid metabolism than simple medication.

Acupuncture Points ; Adult ; Body Mass Index ; Electroacupuncture ; Female ; Humans ; Hypolipidemic Agents ; administration & dosage ; Lipid Metabolism ; Lipids ; blood ; Male ; Metabolic Syndrome ; drug therapy ; metabolism ; therapy ; Middle Aged

OBJECTIVEDuchenne muscular dystrophy (DMD) is an X-linked recessive disease caused by dystrophin gene mutations; 55%-65% of these pathogenic mutations are large deletion and duplication mutations that can be detected by multiplexed polymerase chain reaction. However, finding the remaining micro-mutations (substitutions, deletions or insertions of one or several nucleotides) cannot be achieved in this way. The aim of the present study was to detect mutations of the dystrophin gene in individuals with Duchenne muscular dystrophy (DMD) by denaturing high-performance liquid chromatography (DHPLC) and to establish a rapid and sensitive screening platform for micro-mutations leading to DMD.

METHODSTwenty patients negative for large deletions in the dystrophin gene by multiplex PCR were selected for further screening by DHPLC and 20 normal male without DMD family history as the control cohort. Dystrophin exons and their flanking sequences were individually amplified by genomic PCR and the amplicons showing abnormal DHPLC profile were directly sequenced to identify the position and the type of the mutations.

RESULTSAfter screening 68 exons covering the two deletion hotspots and 3'UTR region, four pathogenic mutations, including c.6808_6811del TTAA, c.4959_4960insA, c.8656C > T and c.8608C > T, were found in four DMD patients. Moreover, c.6808_6811del TTAA, c.4959_4960ins and c.8656C > T have not been reported previously. The first two frameshift mutations were predicted to produce premature stop codons, p.Leu2270MetfsX9 and p.Ser1654LysfsX5, respectively. The remaining two were nonsense mutations, leading to p.R2886X and p.R2870X, respectively.

CONCLUSIONThree novel and one recurrent dystrophin mutations have been identified in Chinese DMD patients. This study has demonstrated that DHPLC is an effective screening method for micro-mutation associated with DMD.

Chromatography, High Pressure Liquid ; methods ; trends ; DNA Mutational Analysis ; Dystrophin ; genetics ; Humans ; Infant ; Male ; Muscular Dystrophy, Duchenne ; genetics ; Mutation ; Sequence Deletion

OBJECTIVETo explore the association between the short tandem repeat polymorphism of exon 5 of MICA gene (MICA-STR) and nasopharyngeal carcinoma (NPC) in a southern Chinese population.

METHODSOne hundred and twenty-seven consecutive NPC patients and 112 randomly selected normal controls residing in southern China mainland were analyzed for MICA-STR allelic variation and MICA gene deletion by fluorescent polymerase chain reaction-gene scanning and polymerase chain reaction-sequence specific priming.

RESULTSMICA*A9 was observed at significantly higher frequency in the NPC patient group than in the control group (relative risk = 2.524, P = 0.001,Pc = 0.006); whereas MICA*A5.1 was present at significantly lower frequency in the NPC patient group than in the control group (RR = 0.418, P = 0.0004, Pc = 0.0026). Further analysis revealed that MICA*A9 was over-represented in male NPC patients, compared with male controls (RR = 3.23, P = 0.00095, Pc = 0.006); whereas MICA*A5.1 was present at significantly lower frequency in male NPC patients, compared with male controls (RR = 0.372, P = 0.0007, Pc = 0.004). None of the MICA-STR variants showed statistically significant frequency difference between female NPC patients and female controls (Pc > 0.05).

CONCLUSIONMICA-STR polymorphism is associated with NPC, and MICA*A9 is a genetic susceptibility marker of male individuals for NPC in a southern Chinese population.

Adult ; Asian Continental Ancestry Group ; genetics ; China ; Exons ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Histocompatibility Antigens Class I ; genetics ; Humans ; Male ; Microsatellite Repeats ; genetics ; Middle Aged ; Nasopharyngeal Neoplasms ; ethnology ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics