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 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 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

To study the impact of the enterovirus 71(EV71) on the nuclear transport mechanism,The pGFP-NLS vector with nuclear location signal(NLS) was constructed, RD cells transfected by the pGFP-NLS vector were inoculated with the EV71 or cotransfected by EV71-2A vector. The results showed that GFP protein with NLS was expressed in the cytoplasm due to the inhibition of nuclear transport. In order to further study the mechanism of the EV71 to prevent nuclear transport,Nup62 was detected by Western blotting after RD cells were infected with EV71 or transfected by EV71-2A vector. The results showed that decreased expression of Nup62 could be detected after infection with EV71 and transfection by EV71-2A vector. This study demonstrates that the cleavage of Nup62 by EV71 2A protease may be the mechanism of nuclear transport inhibition.
Active Transport, Cell Nucleus ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Enterovirus A, Human ; enzymology ; genetics ; metabolism ; Enterovirus Infections ; virology ; Gene Expression Regulation, Viral ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Membrane Glycoproteins ; metabolism ; Nuclear Localization Signals ; metabolism ; Nuclear Pore Complex Proteins ; metabolism ; Peptide Hydrolases ; metabolism ; Recombinant Fusion Proteins ; metabolism ; Transfection

OBJECTIVETo study the effect of GMCSF-absence on the rate of wound healing and neovascularization during wound repair.

METHODS30 wild type (WT) mice and 30 GMCSF- absence mice (GMCSF-KO) were obtained. They were received full thickness skin wound (0.8 cm x 0.8 cm) in each side of midline after deeply anesthesia. In the different post-injury time points, the wound sites were digitally photographed to calculate the percentage of wound closure by using computer image analyses software. The wound specimens were also obtained dynamically for immunohistological analysis of CD31 at wound site.

RESULTSThe analysis of the wound closure showed that wound healing in GMCSF-KO mice was delayed significantly comparing with that in WT mice from the day 3 post-wounding. At days 7 and 10 after wounding significantly more numbers of blood vessels were formed in the WT controls compared to the GMCSF-KO mice.

CONCLUSIONSGMCSF-absence inhibits neovascularization during wound repair and leads to the delay of wound healing.

Animals ; Disease Models, Animal ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; deficiency ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Monocytes ; Neovascularization, Physiologic ; Skin ; blood supply ; injuries ; Wound Healing

OBJECTIVETo study the changes in pelvic floor morphology and relationships of the pelvic organs in patients with outlet obstructive constipation (OOC).

METHODSFifty-nine OOC patients and 12 healthy volunteers were examined by simultaneous pelvic four-contrast defecography, including pelvicography, vaginal opacification, voiding cystography and defecography. The levels of perineum, peritoneum and bladder, and anorectal angle were measured and recorded.

RESULTSAmong the 59 OOC patients, 46 cases of internal rectal prolapse (IRP), 29 cases of pelvic floor hernia, 7 cases of rectocele (RC), 7 cases of spastic pelvic floor syndrome (SPFS), 5 cases of descending perineum syndrome were diagnosed by pelvic four-contrast defecography. Six cystoceles and 10 uterine prolapses were also found by pelvic four-contrast defecography. Compared with healthy volunteers, OOC patients had significantly large anorectal angles during defecation (P<0.05), abnormality descending of perineum during rest and defecation (P<0.05, P<0.01), and a deep pouch of Douglas during defecation (P<0.01). OOC patients combined with urinary system symptoms had an abnormal descent of the bladder during rest and defecation (P<0.05, P<0.01).

CONCLUSIONPelvic four-contrast defecography is an effective method for the diagnosis of IRP, RC, pelvic floor hernia, peritoneocele, cystocele and uterine prolapse, and is helpful in the selection of proper surgical procedures for the treatment of OOC.

Adult ; Aged ; Case-Control Studies ; Constipation ; diagnostic imaging ; etiology ; Defecography ; methods ; Female ; Humans ; Intestinal Obstruction ; complications ; Male ; Middle Aged ; Pelvis ; diagnostic imaging

OBJECTIVETo evaluate the methods and efficacy of atrial septal defect (ASD) occlusion through right chest incision.

METHODSThe clinical data of 21 patients with ASD from July 2004 to May 2005 were analyzed retrospectively. Eight patients were male and 13 patients were female, aged from 1 to 70 years old, with the median age of 21 years old. The diameter of ASD was from 8 to 40 mm. All the 21 patients were under general anaesthesia. A 2 to 3 cm incision was made in the 4th intercostals of right side of sternum. With the help of transesophageal or normal transthoracic echocardiography, the occluder was released using monotube unit.

RESULTSAll cases were occluded successfully without death. The types of the occluder were from 14 to 46 mm. None failed and had to choose extracorporeal circulation operations. No transfusion and no serious complication such as the occluder dislocated occurred. And no evident of atrial shunt was found when in review echo.

CONCLUSIONSThe ASD occlusion through right chest minimal incision is safe, credit, minimal invasive and worth to use widely.

Adolescent ; Adult ; Aged ; Cardiac Catheterization ; methods ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Heart Septal Defects, Atrial ; surgery ; Humans ; Infant ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Retrospective Studies ; Treatment Outcome

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