MP-Rep fusion gene which size is about 1000bp was constructed by recombinant PCR technique.In order to construct fusion gene,Tobacco mosaic virus(TMV) partial movement protein gene(MP) and Cucumber mosaic virus(CMV) partial replicase gene(rep) were ligated.Two copies of MP-Rep fusion gene were ligated with soybean intron in inverted repeat manner,the recombinant fragments were then inserted into binary vector pBIN438 under the control of 35S promoter.Recombinant clone pBIN438-MP-Rep(i/r) which contained two different virus derived genes was constructed.Recombinant clone pBIN438-MP-Rep(i/r) in accord with expected design was certified by restriction endonuclease enzymes digestion and PCR analysis.This approach provides a basis for Broad-spectrum plant virus resistance mediated by RNA Silencing.

OBJECTIVETo explore the relationship between Helicobacter pylori (Hp) infection and iron status using serum ferritin (SF) as a marker for total iron and to identify the related factors of iron nutritional status among preschool children.

METHODSBy cluster sampling, we recruited 475 preschool children aged 2 to 7 years. A structured questionnaire and diet form were sent to the parents of these children to obtain related information about the socioeconomic level and dietary intakes. After collecting blood samples, the following indexes were measured. Hp IgG antibodies were measured with a dot enzyme-linked immunoassay; hemoglobin, Hct, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red blood cell distribution width index (RDW) with automatic Complete Blood Count; SF with an immunoradiometric assay. Stool Hp antigen and occult bleeding were measured with ELISA among individuals who were Hp seropositive. Hp status was defined as positive when both serum and stool antigen tests were positive, Hp status was defined as negative when serum antigen test was negative; 24-hour weighting and recording methods were used to dietary survey for three days in May and December 2001, respectively, dietary intakes including energy, protein and micronutrient were calculated using nursery school nutrition software and evaluated by Chinese Dietary Reference Intakes (DRIs). Mann-Whitney test was used to compare mean ranks of SF in Hp-positive and Hp-negative children. To obtain an adjusted estimate of the impact of Hp infection on SF, a multivariate analysis of covariance was done to evaluate the different level of SF between Hp infected and non-infected status. The relationship between iron deficiency and gender, age, socioeconomic condition, iron intake, and calcium intake was assessed by univariate analysis. An unconditional multivariate logistic regression analysis was also performed. Iron deficiency status was dichotomized and placed as the dependent variable. Hp infection status was incorporated together with possible confounding factors as independent variables in a final logistic regression model. All the data were managed by EPI Info 5.01a and analyzed by SAS (Version 6.12).

RESULTSTotally 64 children were diagnosed as Hp-positive and 305 as Hp-negative. Mann-Whitney test and multivariate analysis of covariance both showed that SF concentration was significantly lower in Hp infected individuals than non-infected individuals. Adjusted mean level and 95% confidence interval of SF in infected and non-infected children was 23.62 microg/L (7.13 microg/L-78.26 microg/L), 33.48 microg/L (10.28 microg/L-109.06 microg/L), respectively. The relationship between Hp infection and iron deficiency status persisted in logistic regression analysis after adjusting for possible confounding factors (OR: 7.95; 95% CI 2.56 - 24.67).

CONCLUSIONIron nutritional status was reduced in Hp infected preschool children. Hp infection appears to be an independent risk factor or an added stressor on iron status among preschool children.

Antibodies, Viral ; blood ; Child ; Child, Preschool ; Erythrocyte Indices ; Female ; Ferritins ; blood ; Helicobacter Infections ; blood ; Helicobacter pylori ; immunology ; Hemoglobins ; analysis ; Humans ; Logistic Models ; Male ; Multivariate Analysis ; Nutrition Assessment

Recent studies indicate that beta-amyloid (Abeta) is the key factor to cause neuronal degeneration in Alzheimer's disease (AD). In the present study, we set up an Abeta induced PC12 cell damage modle and studied the protective effect and related mechanisms of T(10), monomer extracted from Chinese herb Tripterygium wilfordii Hook F. PC12 cells were treated with different concentrations of Abeta (5x10(-4), 5x10(-3), 5x10(-2), 5x10(-1), 5, 50 micromol/L) for 48 h, cell viability was detected by MTT conversion. The apoptotic rate of PC12 cells was quantitatively determined using FACS assay. After PC12 cells were treated with 1x10(-11) mol/L T(10) for 48 h and then co-treated with 50 micromol/LAbetafor 48 h, the apoptotic rate and the change in intracellular Ca(2+) concentration of PC12 cells were analyzed by FACS assay and confocal, respectively. It was found that 5 micromol/L Abeta decreased the cell viability to 66.3% and 50 micromol/L Abeta decreased it to 55.1%, significantly different from that of the control group. After treatment with 50 micromol/L Abeta for 48 h, the apoptotic rate of PC12 cells increased obviously. The apoptotic rate was 5.37% in the control group, while after treatment with 0.5, 5 and 50 micromol/L Abeta for 48 h, the apoptotic rate of PC12 cells went up to 10.19%, 8.02% and 16.63%, respectively. At the same time, the concentration of intracellular Ca(2+) increased greatly after treatment with 50 micromol/L Abeta for 48 h. At the concentration of 1x10(-11) mol/L T(10) remarkably inhibited the apoptosis induced by 50 micromol/L Abeta. In the naive group, the apoptotic rate was 4.83%. The apoptotic rate went up to 17.24% after treatment with 50 micromol/L Abeta for 48 h. After co-treatment with 1x10(-11) mol/L T(10) and 50 micromol/L Abeta, the apoptotic rate decreased to 8.91%, significantly different from that of the control group. At the same time, at the concentration of 1x10(-11 )mol/L T(10) remarkably inhibited the increase of intracellular Ca(2+) concentration induced by Abeta. The results indicate that T(10) has obvious protective effect on PC12 cells, which may be related to the inhibition of the cell apoptosis and increment of intracellular Ca(2+) concentration induced by Abeta.
Alzheimer Disease ; pathology ; Amyloid beta-Peptides ; toxicity ; Animals ; Apoptosis ; drug effects ; Calcium ; metabolism ; Diterpenes ; pharmacology ; Epoxy Compounds ; Neuroprotective Agents ; pharmacology ; PC12 Cells ; Peptide Fragments ; toxicity ; Phenanthrenes ; pharmacology ; Rats ; Tripterygium ; chemistry

OBJECTIVETo investigate the effects of Apo A1 and B gene polymorphism on avascular necrosis of the femoral head (ANFH) in north Chinese Han population.

METHODSPolymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) technique was used in samples of 143 cases with documented ANFH and 92 healthy control matched by age and sex individuals selected from north Chinese Han nationality. The studied loci include promoter region (-75bp) and the intron 1 (+83 bp) of Apo A1 gene, Eco RI,Xba I of Apo B gene; polymerase chain reaction was used to study 3'-VNTR of Apo B gene.

RESULTSAt -75 bp in promoter, the frequency of A/A genotype in ANFH group was significantly higher than that in control group (P < 0.01), while the frequency of G/A genotype in ANFH group was significantly lower than that in control group (P < 0.01). No difference was found in the frequency of genotype at +83bp in intron 1 of Apo A1 gene, Eco RI, Xba I and 3'-VNTR loci of Apo B gene.

CONCLUSIONApolipoprotein A1 gene A/A substitution at position -75 in promotor is associated with ANFH, the mutation may be one of the sensitive genes of ANFH, first reported inside and abroad. But no evident relationship was found between gene polymorphism of +75 bp loci of Apo A1 gene, Eco RI loci of Apo B gene, Xba I loci of Apo B gene or 3'-VNTR of Apo B gene and ANFH.

Adult ; Apolipoprotein A-I ; genetics ; Apolipoproteins B ; genetics ; Case-Control Studies ; China ; Electrophoresis ; Female ; Femur Head Necrosis ; genetics ; Gene Frequency ; Genotype ; Humans ; Male ; Polymorphism, Genetic

OBJECTIVETo assess the efficacy and safety of Saw Palmetto Extract Capsules in the treatment of benign prostatic hyperplasia (BPH).

METHODSWe conducted a multi-centered open clinical study on 165 BPH patients treated with Saw Palmetto Extract Capsules at a dose of 160 mg qd for 12 weeks. At the baseline and after 6 and 12 weeks of medication, we compared the International Prostate Symptom Scores (IPSS), prostate volume, postvoid residual urine volume, urinary flow rate, quality of life scores (QOL), and adverse events between the two groups of patients.

RESULTSCompared with the baseline, both IPSS and QOL were improved after 6 weeks of medication, and at 12 weeks, significant improvement was found in IPSS, QOL, urinary flow rate, and postvoid residual urine. Mild stomachache occurred in 1 case, which necessitated no treatment.

CONCLUSIONSaw Palmetto Extract Capsules were safe and effective for the treatment of BPH.

Capsules ; Humans ; Male ; Plant Extracts ; adverse effects ; therapeutic use ; Prostatic Hyperplasia ; drug therapy ; Quality of Life

OBJECTIVETo study the relationship between two potential functional polymorphisms in exon 2 of the p73 gene and the susceptibility of lung cancer.

METHODSGenotypes were determined by polymerase chain reaction-single stand conformation polymorphism (PCR-SSCP) method in 425 histologically-confirmed lung cancer cases and 588 cancer-free controls, frequency-matched by age and sex.

RESULTSThe two polymorphisms were in complete linkage disequilibrium and the frequencies of variant p73 AT haplotype (A4T14) were less commonly seen in the cases (0.225) than in the controls (0.287) (P = 0.0018). Compared with the p73 GC/GC homozygotes, both the AT/AT variant homozygotes and GC/AT heterozygotes were associated with a significantly decreased risk [adjusted odds ratio (OR) = 0.45, 95% confidence interval (CI) = 0.26 - 0.80 and OR = 0.70, 95% CI = 0.53-0.92, respectively].

CONCLUSIONThese results suggested that this p73 dinucleotide polymorphism might have had a role to play in the susceptibility of lung cancer.

5' Untranslated Regions ; genetics ; Adenocarcinoma ; genetics ; Adult ; Aged ; Carcinoma, Small Cell ; genetics ; DNA-Binding Proteins ; genetics ; Exons ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; physiology ; Genetic Predisposition to Disease ; genetics ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Nuclear Proteins ; genetics ; Polymorphism, Genetic ; Tumor Protein p73 ; Tumor Suppressor Proteins ; genetics

OBJECTIVETo study the surgical reconstruction of unilateral Craniofacial Atrophy and Hypoplasia.

METHODSAccording to the etiological factors and severity of the facial deformities, different methods are employed, including bone framework reconstruction, soft tissue transplantation, orthognathic surgery.

RESULTSFrom September 1998 to August 2004, 42 cases were treated, Include: Hemifacial Microsomia 22 cases, Hemifacial Atrophy 16 cases, unilateral facial hypoplasia due to radiation 4 cases. Miniplate and transplants extrusion occurred on 2 post radiation patients due to poor soft tissue coverage, infection occurred on 1 patient after mandibular ramus reconstruction using autogenous rib and contralateral mandibular outer cortex. The leaving patients recovered well and the facial asymmetry were improved greatly.

CONCLUSIONSFacial asymmetry due to unilateral Craniofacial Atrophy or Hypoplasia is a common and complex condition for surgical management, The surgical plan should be delicated made individually according to the severity of the soft tissue and the underlying bone framework.

Adolescent ; Adult ; Craniofacial Abnormalities ; surgery ; Facial Asymmetry ; surgery ; Facial Hemiatrophy ; surgery ; Female ; Humans ; Male ; Reconstructive Surgical Procedures ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo study the association between the single nucleotide polymorphisms (SNPs) in FXYD6 gene and schizophrenia in a family-trios population.

METHODSSix SNPs (rs10790212, rs11544201, rs555577, rs1815774, rs4938446 and rs497768) in the FXYD6 gene were genotyped by allele-specific PCR method in 101 nuclear families, and transmission disequilibrium test (TDT) was performed.

RESULTSSNPs rs10790212 and rs11544201 showed significant association with schizophrenia (P<0.05). Furthermore, significant association of schizophrenia with the haplotype rs10790212-rs11544201 was found (P<0.05).

CONCLUSIONFXYD6 gene might play an important role in schizophrenia susceptibility and functional analysis of FXYD6 are needed.

Adolescent ; Adult ; Alleles ; Female ; Genetic Predisposition to Disease ; Haplotypes ; Humans ; Ion Channels ; genetics ; Linkage Disequilibrium ; genetics ; Male ; Polymorphism, Single Nucleotide ; genetics ; Schizophrenia ; genetics ; Young Adult

OBJECTIVETo investigate the main risk factors for postoperative severe complications, and establish Logistic regression model to predict severe complications in gastric cancer following gastrectomy.

METHODSThe data of 1728 gastric cancer patients underwent gastrectomy between June 2001 and June 2007 were analyzed retrospectively. Logistic regression analysis was used to investigate the risk factors for postoperative severe complications in those patients.

RESULTSPostoperative severe complications were associated with extent of lymph node dissection (D(2)(+)-D(3)), chronic obstructive pulmonary disease (COPD), invasion to the adjacent organ, combined organ resection, extent of lymph node dissection (D(2)), diabetes mellitus (DM), TNM staging IV, heart diseases, malnutrition, surgeon's operative volume, operative time, blood loss and age. The Logistic regression model was P = 1/[1+e((14.806-2.523X1-1.792X2-1.558X3-1.551X4-1.270X5-1.150X6-1.101X7-0.981X8-0.817X9-0.657X10-0.578X11-0.542X12-0.309X13))]. A testing sample showed that the accuracy, sensitivity and specificity of the Logistic model were 72.5%, 70.0% and 75.0%, respectively.

CONCLUSIONSThe extent of nodal dissection (D(2)(+)-D(3)), COPD, invasion to the adjacent organ, combined organ resection, extent of nodal dissection (D(2)), diabetes mellitus, TNM staging IV, heart diseases, malnutrition, surgeon's operative volume, operative time, blood loss and age are the independent risk factors associated with severe complications in gastric cancer post gastrectomy. The Logistic regression model based on these factors is reliable in predicting the severe complications.

Adult ; Aged ; Aged, 80 and over ; Female ; Gastrectomy ; adverse effects ; Humans ; Logistic Models ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; surgery

Objective:To evaluate effects of Bushen Jianpi Recipe ( BSJP) on restoring of T cell exhaustion induced by persistent tuberculosis antigens in mice.Methods:The C57BL/6 mice were primed with BCG(5×106 CFU)and boosted with 20μg of Mycobacterium bovis BCG Purified Protein Derivative ( PPD ) once a week and lasted for 16 times to induce T cell exhaustion that happened in late stage of tuberculosis .BSJP was used to treat the mice with immune exhaustion for 3 weeks.Flow Cytometry was used to detect the numbers of CD 4+and CD8+T cells and PD-1 expression.ELISA was used to detect IL-2 and IFN-γsecretion.Bacteria load in lung tissue following BCG challeng was used to detect the protective capability of mice models .Results: Compared with the transient antigen stimulation group,the persistent antigen stimulation group had lower level of IFN-γand IL-2 secretions(P<0.01).The numbers of CD4+and CD8+T cells decreased and the level of inhibitory receptor PD-1 got higher(P<0.01).The protective efficacy against BCG challenging decreased ( P<0.01 ) .After treatment with BSJP , the production of cytokines IFN-γand IL-2 increased ( P<0.01 );the numbers of CD4+and CD8+T cells increased and PD-1 expression level on CD4+T cells decreased(P<0.01);the protective efficacy against BCG challenge increased (P<0.01).The results showed that BSJP can reduce the PD-1 expression,improve the IL-2 and IFN-γsecretion,and increase the protection against BCG challenge .Conclusion:Bushen Jianpi Recipe can restore T cell exhaustion induced by persistent tuberculosis antigen stimulation in mice , which gives a hope to overcome T cell exhaustion in tuberculosis with Chinese herbal in clinic.