M2e gene of three copies for H5N1 subtype AIV was synthesized and fused with human mycobacterium tuberculosis hsp70 gene.The fused gene was cloned into the prokaryotic expression vector to get pET-3M2e and pET-3M2e-hsp70.Recombinant protein r3M2e and r3M2e-hsp70 were successfully expressed induced with IPTG and purified with Ni2+-NTA collumn.Following that,the immunity of the recombinant protein was analysized with Western blot.20-day-old AIV non-immunized chickens were vaccined with r3M2e and r3M2e hsp70,at the same time,Trx and KLH-M2e inoculated chickens were served as vector and positive controls.Two weeks after the primary vaccination,every group was boosted with the same vaccine as in the primary vaccination.The humoral immunity of the vaccined chickens was evaluated with antibody detection against M2e,cytopathic suppression test,and indirect fluorescence assay.The cellular immunity was estimated according to lymphocyte subtype analysis with flow cytometry and M2e specific cytokine detection.Four weeks after the boost vaccine,all groups were challenged with 100EID50 AIV of H9N2 subtype,and the virus from swabs was detected with Real-time PCR.Results indicated that r3M2e hsp70 vaccined chicken developed the better humoral and cellular immune response,also,made a better performance compared with r3M2e vaccined group in virus challenge.

Objective To investigate the current situation of the prevention and contml of iodine deficiency disorders(IDD)in Hubei Province,SO as to provide a policy.making basis for controlling work of IDD. Methods Using the method of proportional population sampling(PPS),30 eounties were seiected in Hubei Province.In each selected county,1 primary school was chosen.In every primary school,40 pupil8 aged 8～10 years were selected to examine thyroid size,intelligent quotient(IQ),and salt iodine contents at their home.In the selected pupils,2 boys and 2 girls were chosen to determine their urinary iodine contents in each age grouD.Twenty pupils in the above school and 5 housewives ileal"to this school were tested in health education questionnaire. Results The median of salt iodine was 30.1 mg/kg,and the rate of comsuming qualified iodized salt was 96.2% (1154/1200).The rates ofchild goiter were 6.5%(78/1200)by palpation and 4.1%(49/1200)by B ultrasound.The median of urinary iodine Was 358.4μg/L and mean of the IQ was 105.3±14.4.The rate of qualified scores of both students and housewives Was 25.5%(153/600)、90.7%(136/150).Conclusions The current Bituation of iedine nutrition is good.The goal of eliminating IDD has been achieved in Hubei Province.

OBJECTIVETo summarize clinical application result of the thoracodorsal artery perforator pedicled flap for repair of soft tissue defects on the ipsilateral upper limb.

METHODSFrom September 2003 to May 2007, 8 patients (6 males and 2 females) with soft tissue defects on the ipsilateral upper limb underwent reconstruction with the thoracodorsal artery perforator pedicled flap. The age of patients was from 16- to 45-years-old with an average of 32 years. Of them, the recipient sites of 5 cases were located on the arm region, 3 cases on the forearm.

RESULTSThe minor superficial infection of 1 case occurred on the recipient site after operation and the wound gradually healed by daily change dressings. All the flaps had survived completely and the postoperative course was uneventful with satisfactory clinical results. Follow-up period ranged for 9-38 months after operation (mean, 19 months). There was no remarkable donor site morbidity. All cases had good appearance on recipient site.

CONCLUSIONThe thoracodorsal artery perforator pedicled flap is thin and suitable for repair of soft tissue defect on the ipsilateral upper limb.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Soft Tissue Injuries ; physiopathology ; surgery ; Surgical Flaps ; blood supply ; Thoracic Arteries ; surgery ; Upper Extremity ; blood supply ; surgery ; Young Adult

OBJECTIVETo study the effect and mechanism of Ganbi decoction (GBD) in treating patients with antituberculotic agent caused liver injury (ATB-LI).

METHODSOne hundred and twenty-eight patients with ATB-LI were randomly assigned to the treated group (n = 66) and the control group (n = 62) with the envelop method. Meanwhile, 60 healthy persons were selected as the healthy control group. The treated group was treated by GBD one dose every day with the constituents modified depending on patients' symptoms, and the control group was treated with glucuronolactone tablets and inosine injection. One week was taken as one treatment course. The changes of clinical syndromes, physical signs, T-lymphycyte sub-groups and serum level of nitric oxide (NO) were observed before and after treatment and the recovery time of liver function was recorded. The outcome was compared with that in the healthy control group.

RESULTSIn the treated group, 28 patients (42.4%) were cured, 30 (45.5%) improved and 8 (12.1%) ineffectively cured, the total effective rate being 87.9% (58/66). In the control group, 17 patients (27.4%) were cured, 24 (38.7%) improved, and 21 (33.9%) ineffectively cured, the total effective rate being 66.1% (41/62). The total effective rate in the treated group was significantly higher than that in the control group (P < 0.05). Liver function was improved in both groups, recovery time in the treated group was 12.0 +/- 7.0 days, which was significantly shorter than that in the control group (16.0 +/- 8.0 days), showing significant difference between the two groups (P < 0.05). The levels of CD3, CD4 and CD8 were significantly higher and level of NO significantly lower in the two groups of patients than those in the healthy control group (P < 0.05), but these parameters were improved more significantly in the treated group after treatment, when compared with those before treatment or with those in the control group, all showing significant difference (P < 0.05).

CONCLUSIONGBD could prevent ATB-LI, and its mechanism could be by way of reducing NO production induced by endotoxin of macrophage and stimulating the proliferation of T-lymphycyte to elevate immunity.

Adult ; Aged ; Antitubercular Agents ; adverse effects ; Chemical and Drug Induced Liver Injury ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glucuronates ; therapeutic use ; Humans ; Inosine ; therapeutic use ; Liver Diseases ; drug therapy ; Liver Function Tests ; Male ; Middle Aged ; Nitric Oxide ; blood ; T-Lymphocyte Subsets ; Treatment Outcome

OBJECTIVETo investigate the impact of PET/CT on the clinical staging, target volume delineation and precise radiotherapy (PAR) planning for patients with non-small cell lung cancer (NSCLC).

METHODSPET/CT scanning was performed in 58 histologically proven NSCLC patients for radical radiotherapy or surgery. The clinical staging of all patients was determined by PET/CT according to 1997 World Health Organization (WHO) staging system. The gross tumor volume (GTV) was delineated and three-dimensional conformal radiotherapy (3D-CRT) planning was established with identical parameters based on CT image and PET/CT fused image, respectively. The indexes including volume of GTV (V(GTV)), percentage of the total lung volume which received more than 20 Gy (V(20)), mean lung dose (MLD), tumor control probability (TCP), normal tissue complication probability (NTCP), and dose to spinal cord (Ds) were selected and evaluated. The quality of the two plans and the impact of PET/CT on PAR planning was compare and analyzed.

RESULTS1. PET/CT image results changed the clinical stages in 21 of 58 (36.2%) patients with 14 upstaged and 7 downstaged, therefore, the management decisions were modified in 16 (27.6%) patients. 2. Among 32 patients who underwent surgery, PET/CT staging result was consistent with pathologic staging in 29 with one false negative and 2 false positive in lymph node staging. The sensitivity of PET/CT was 96.9% and accuracy 90.6%. 3. The differences of indexes including V(GTV) (P = 0.004), V(20) (P = 0.000) and MLD (P = 0.004) between the two radiotherapy plannings were statistically significant, whereas, the Ds, TCP and NTCP (left lung, right lung, skin and spinal cord) was not.

CONCLUSION1. The impact PET/CT on clinical staging of NSCLC and PAR planning was remarkable. 2. PET/CT is more consistent with pathology in staging than CT and, therefore, is an important compensatory staging measure. 3. Compared with CT, PET/CT can reduce the V(GTV) in patients with atelectasis and obstructive pneumonitis when contouring the target volume, so can provide better protection for normal surrounding lung tissue. On the other hand, PET/CT is more sensitive in detecting mediastinal lymph node metastasis than CT, and the V(GTV) can be more precise and guaranteed. 4. Radiopneumonitis may be more effectively prevented because of significant decrease in V(20) and MLD by more precise planning based on PET/CT results. 5. PET/CT not only can provide satisfactory Ds, TCP and NTCP within clinical demand, but also more precise delineation of the radiation target volume and precise radiotherapy planning for NSCLC.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; diagnostic imaging ; pathology ; radiotherapy ; surgery ; Female ; Humans ; Lung Neoplasms ; diagnostic imaging ; pathology ; radiotherapy ; surgery ; Male ; Middle Aged ; Neoplasm Staging ; Positron-Emission Tomography ; methods ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Conformal ; methods ; Tomography, X-Ray Computed

OBJECTIVETo develop a high-throughput screening assay for Farnesoid X receptor (FXR) agonists based on mammalian one-hybrid system (a chimera receptor gene system) for the purpose of identifying new lead compounds for dyslipidaemia drug from the chemical library.

METHODScDNA encoding the human FXR ligand binding domain (LBD) was amplified by RT-PCR from a human liver total mRNA and fused to the DNA binding domain (DBD) of yeast GAL4 of pBIND to construct a GAL4-FXR (LBD) chimera expression plasmid. Five copies of the GAL4 DNA binding site were synthesized and inserted into upstream of the SV40 promoter of pGL3-promoter vector to construct a reporter plasmid pG5-SV40 Luc. The assay was developed by transient co-transfection with pG5-SV40 Luc reporter plasmid and pBIND-FXR-LBD (189-472) chimera expression plasmid.

RESULTSAfter optimization, CDCA, a FXR natural agonist, could induce expression of the luciferase gene in a dose-dependent manner, and had a signal/noise ratio of 10 and Z' factor value of 0.65.

CONCLUSIONA stable and sensitive cell-based high-throughput screening model can be used in high-throughput screening for FXR agonists from the synthetic and natural compound library.

Base Sequence ; Cell Line ; DNA Primers ; DNA, Complementary ; DNA-Binding Proteins ; agonists ; chemistry ; genetics ; Humans ; Hypolipidemic Agents ; analysis ; Plasmids ; Receptors, Cytoplasmic and Nuclear ; agonists ; chemistry ; genetics ; Reproducibility of Results ; Transcription Factors ; agonists ; chemistry ; genetics ; Transfection

OBJECTIVETo study the distribution, frequency and structure of variable number of tandem repeats (VNTR) in 3' region of apoB gene in Chinese population.

METHODSGenomic DNA was extracted from peripheral blood obtained under consent from randomly-chosen 522 individuals who came to the hospital for physical examination, and used to screen for polymorphisms of 3' VNTR of the apoB gene by employing polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), gradient polyacrylamide gel electrophoresis, cloning and sequencing.

RESULTSSixteen types of alleles of apoB 3oVNTR were identified, among which heterozygotes were more than homozygotes. The biggest allele is HVE58, and the smallest one is HVE22. HVE34 had the highest frequency (40.4%), followed by HVE32 (34.7%). This showed significant difference from the allelic distribution of other populations (Caucasian and Swedish). Through sequencing of 60 alleles, a new isomer (Y-A=ATAATTAAATATTT) and four new types of alleles were found.

CONCLUSIONThe Chinese population we studied had a higher frequency of small alleles and showed a difference in allelic structure and frequency distribution from European and American in this populations.

Adult ; Aged ; Aged, 80 and over ; Alleles ; Apolipoproteins B ; genetics ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Male ; Middle Aged ; Minisatellite Repeats ; genetics ; Polymerase Chain Reaction

OBJECTIVETo study the pattern of lymph node metastasis of thoracic esophageal squamous cell carcinoma (ESCC) after esophagectomy and its impact on the clinical target volume (CTV) delineation in radiotherapy fpr thoracic ESCC.

METHODSThe pattern of lymph node metastasis was retrospectively analyzed in 1077 patients with primary thoracic ESCC. All patients received esophagectomy with two- or three-field lymphadenectomy. The clinicopathologic factors related to lymph node metastasis were then analyzed using logistic regression analysis.

RESULTSThe rates of cervical, upper mediastinal, middle mediastinal, lower mediastinal and abdominal cavity lymph node metastasis were 16.7%, 33.3%, 11.1%, 5.6% and 5.6%, respectively. The rates of those node metastasis in the middle thoracic ESCC were 4.0%, 3.8%, 28.5%, 7.1% and 17.1%, respectively, and the rates of those node metastasis in the lower thoracic ESCC were 1.5%, 3.0%, 22.7%, 37.0% and 33.2%, respectively. The depth of tumor invasion, histologic differentiation and the length of tumor were showed to be statistically most significant risk factors of lymph node metastasis of ESCC (P < 0.001).

CONCLUSIONThe depth of tumor invasion, histologic differentiation, and length of tumor were closely correlated with lymph node metastasis of ESCC. All these factors and tumor location should be considered comprehensively when designing the target volume for radiotherapy.

Aged ; Carcinoma, Squamous Cell ; pathology ; radiotherapy ; Esophageal Neoplasms ; pathology ; radiotherapy ; Esophagectomy ; Female ; Humans ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Proportional Hazards Models ; Radiotherapy, Conformal ; methods ; Retrospective Studies ; Risk Factors

BACKGROUNDAtrial fibrillation (AF) is accompanied by atrial structural remodeling. Calpain activity is induced during AF. To test a causal relationship between calpain activation and atrial structural changes, N-acetyl-Leu-Leu-Met (ALLM), a calpain inhibitor, was utilized in a canine AF model.

METHODSFifteen dogs were randomly divided into 3 groups: sham-operated group, control group and calpain inhibitor group; each with 5 dogs. Sustained AF was induced by rapid right atrium pacing at 600 beats per minute for 3 weeks. ALLM was administered at a dosage of 1.0 mg x kg(-1) x d(-1) in the calpain inhibitor group. Three weeks later, the proteolysis, protein expression of TnT and myosin, calpain I localization and expression and structural changes were examined in left atrial free walls, right atrial free walls and the interatrial septum respectively. Atrial size and contractile function were also measured by echocardiography.

RESULTSLong-term rapid atrial pacing induced marked structural changes such as enlarged atrial volume, myolysis, degradation of TnT and myosin, accumulation of glycogen and changes in mitochondrial shape and size, which were paralleled by an increase in calpain activity. The positive correlation between calpain activity and the degree of myolysis (r(s) = 0.90 961, P < 0.0001) was demonstrated. In addition to structural abnormalities, pacing-induced atrial contractile dysfunction was observed in this study. The pacing-induced atrial structural alterations and loss of contractility were partially prevented by the calpain inhibitor ALLM.

CONCLUSIONSActivation of calpain represents key features in the progression towards overt structural remodeling. Calpain inhibitor, ALLM, suppressed the increased calpain activity and reversed structural remodeling caused by sustained atrial fibrillation in the present model. Calpain inhibition may therefore provide a possibility for therapeutic intervention in AF.

Animals ; Atrial Fibrillation ; pathology ; Calpain ; antagonists & inhibitors ; Cysteine Proteinase Inhibitors ; pharmacology ; Disease Models, Animal ; Dogs ; Heart Atria ; pathology ; ultrastructure ; Myosins ; analysis ; Troponin T ; analysis

OBJECTIVETo investigate the clinical epidemiological features. of blast injury of lungs caused by gas explosion.

METHODSA retrospective analysis of 132 cases of blast injury of lungs caused by gas explosion injuries in our hospital from 1990 to 2010 were made, including the information of lung injury, associated injuries, treatment and mortality.

RESULTS56 patients were conscious on admission, during 1 ∼ 5 days, 30 patients got cough, hemoptysis and other pulmonary symptoms, 7 patients had hemothorax, 11 cases combined with pneumothorax, 86 cases of patients with burns, 96 cases with pulmonary aspiration compound injury, 41 cases with abdominal organs blast injuries, 76 cases with pulmonary infection, 31 cases with shock, Seven patients died of respiratory failure, two patients died of respiratory failure.

CONCLUSIONBlast injury of lung featured with serious internal injuries, combined with many injuries, The incidence of compound injury was high, and it was difficult to diagnose. Chest X-ray were varied. The mortality was high, and the predominant causes of death were respiratory failure, shock, sepsis. Most patients with blast injury of lung need tracheotomy and mechanical ventilation. Timely tracheotomy, mechanical ventilation were the major prognostic measures.

Adult ; Blast Injuries ; epidemiology ; etiology ; Coal Mining ; Explosions ; Gases ; Humans ; Lung Injury ; epidemiology ; etiology ; Male ; Retrospective Studies ; Young Adult