OBJECTIVE: To construct a plant effective expression vector driven by a fruit specific promoter for the expression of hepatitis B virus surface antigen (HBsAg), to further improve the expression of exogenous gene in plant, and to prepare for the development of an effective anti-hepatitis vaccine. METHODS: Tomato fruit-specific promoters' gene 2A12 and E8 were respectively introduced to pBPFOmega7 to form pB2A12 and pBE8. The DNA fragment containing HBsAg-s gene from plasmid YEP-HBs was inserted respectively into pB2A12 and pBE8 to form pB2A12-HBs and pBE8-HBs. The fragment containing "p35S+2A12+Omega+HBsAg-s+Tnos" of the pB2A12-HBs was sub-cloned into plasmid pCAMBIA1301 to yield the reconstructed plant binary expression plasmid pCAM2A12-HBs, and the fragment containing "p35S+E8+Omega+HBsAg-s+Tnos" of the pBE8-HBs was sub-cloned into plasmid pCAMBIA1301 to yield the plasmid pCAME8-HBs. The inserted gene HBsAg and fruit-specific promoters in the reconstructed plant binary expression vectors were confirmed by sequencing. Then, pCAM2A12-HBs and pCAME8-HBs were directly introduced into Agrobacterium tumefaciens strain EHA105. RESULTS: Digestion with restriction enzymes proved that all recombinant vectors had the inserts with expected length of the target fragments, and the sequencing results were confirmed correct. CONCLUSION: In this study, plant expression vector containing HBsAg gene driven by fruit specific promoter and CaMV35s promoter was successfully constructed.

Objective To study the possibility of targeting endothelial cells with the 131I labeled monoclonal antibody (mAb) for the treatment of hepatocarcinoma. Methods An animal model of human hepatocarcinoma in nude mice was reproduced, and tumor inhibiting activity of the 131I labeled mAb was tested. 30 nude mice with hepatocarcinoma were randomly divided into 3 groups: in group A the mice were treated with mAb 200?g/200?l twice a week; group B mice were treated with the 131I labeled mAb 200?g/200?l twice a week; control group mice were given normal saline in equal volume. Tumor growth in mice was observed. After the mice were sacrificed, the tumor was histologically examined and the intra-tumor microvessel density (TMVD) recorded. Results The tumor growth inhibition effect in mice treated with mAb was 74.55%. This effect was enhanced when treated with the 131I labeled mAb, as evidenced by an increase of tumor growth inhibition rate to 86.36%. Pathologically, Massive necrosis of tumor cells around the degenerated vessels was observed in the mAb treated mice. TMVD was significantly lower in the mAb treated mice than that in the untreated mice (P

Objective To evaluate the efficacy of internal fixation with or without fusion in the treatment of thoracolumbar burst fractures.Methods Clinical controlled trails related to the application of pedicle screw instrumentation with or without fusion for thoracolumbar fractures before March,2012were obtained by searching PubMed,Science Direct,Medline and CNKI.Quality evaluation was made on the included literature,from which data were extracted to integrate various rescarch results by using RevMan 5.1.The quantitative data were analyzed based on the effect scale of mean difference (MD) and bilateral 95％ confidence interval (CI).The numeration data were analyzed in the use of effect scale of odds ratio (OR) and bilateral 95％ CI.The merging of some data was manually completed.Results After retrieving,eight English and one Chinese papers of the clinical controlled trials,and two related Meta analysis were obtained.With exclusion of one repetitive research,eight papers were involved in the review.Meta analysis demonstrated that fusion and non-fusion fixation had no significant differences in aspects of correction of kyphotic angle,correction and correction loss of vertebral body height,neurological function improvement,complication rate,and length of hospital stay.While compared with the fusion fixation,non-fusion fixation showed a more serious correction loss of kyphotic angle,a fewer blood loss and a shorter operation time.Conclusions Non-fusion fixation shows the similar efficacy with fusion fixation in the treatment of some thoracolumbar burst fractures pertaining to releasing compression,restoring spinal stability and preventing complications,but it can also significantly decrease operation time and blood loss.Furthermore,non-fusion fixation may markedly improve patients' quality of life since it restores motion of the instrumented segment after removal of implant and decreases the risk of adjacent segmental degeneration.

Objective To establish antibody titre quantitation method by ELISwithoustandard substance .MethodThe tesgroupof non-specifi,negative control ,specifiand total antibodiewere sefodetecting serantibodieby ELIS.Aftelineafitting of the time-absorbance datof early developing ,the fitted slopewere used athe velocity of absorbance changing , which were denoted by ν0 ,νC,νS,νT.Based on thathe ν-value and the concentration ? of determinand were linearelationship while the substrate waexcessive ,the function with parameteof ν-valueforeflecting the multiple proportionof antibodiecon-centrationbetween specifiand negative control groupcould be deduced .The assessmenof specifiIgG antibodiein serof KM mice immunized with fish collagen waused aan instance .ResultThe function C/C= (ν-ν0 )/(νC-ν0 ) could calculate the mul-tiple proportion(titre) of antibodieconcentrationbetween specifiand negative control group.Conclusion The above method of antibody titre quantitation isuitable fosemi-quantitative analysiwithoustandard substance .

Objective To investigate the influence on the ovarian reserved function of Gn-RH injection in the bilateral laparoscopic cys-tectomy. Methods The clinical data of 100 patients with endometriosis undergoing bilateral laparoscopic cystectomy were included in this study. 50 patients treated by routine bilateral laparoscopic cystectomy were set as control group. 50 patients treated by Gn-RH injection addi-tionally during the surgery were set as study group. The operative situation,serum FSH level,FSH/LH,E2 level,ovarian reserved function, pregnancy situation and recurrent rate were compared between two groups. Results The operative duration and electrocautery times in study group was significantly less than those in control group(P<0. 01). The residual ovarian tissue capsule wall thickness in study group was sig-nificantly more than that in control group(P<0. 01). At different time points,the serum FSH level,FSH/LH and E2 level in study group were significantly less than those in control group(P<0. 01,P<0. 05). The incidence of ovarian reserved function damage in study group was significantly less than those in control group(P<0. 05). The pregnancy rate was significantly higher and the time between surgery and pregnancy was significantly shorter in study group than those in control group(P<0. 05,P<0. 01). The recurrent rate in study group was sig-nificantly less than those in control group(P<0. 01). Conclusion There is a influence on ovarian reserved function of bilateral laparoscopic cystectomy. By inject Gn-RH during surgery,the ovarian reserved function could be protected,which increase the pregnancy rate and reduce recurrence.

Objective To compare superiorities of liver malignant tumors underwent regular and irregular hepatectomies through approaching perioperative factors.Methods 1019 consecutive hepatectomies of liver malignant tumor from 1986 to 2009 at Air Force General Hospital and General Hospital of Chinese PLA were investigated retrospectively according to their medical documentation.Results Multivariate analysis showed that liver malignant tumors on which regular hepatectomy and irregular hepatectomy were performed,there was no significant difference in the blood loss,complications,mortality related to operation,hospital stay,and so on.But the operating time of regular hepatectomy was obvious more than that of irregular hepatectomy (P＜ 0.001,OR=1.004).Conclusions Although for liver malignant tumor,regular hepatectomy seems to be superior to irregular hepatectomy based on oncological theory,in clinical practice,there were no significant difference between the perioperative risk of regular hepatectomy and that of irregular hepatectomy.

Objective To investigate the construction and implementation of the concept of precise sequential therapy for primary liver cancer.Methods The clinical data of 207 patients with resectable liver cancer who were admitted to the Air Force General Hospital from May 2005 to June 2010 were retrospectively analyzed.Of all the patients,81 received conventional therapy (conventional therapy group),and the other 126 patients received precise sequential therapy (precise sequential therapy group).The conditions of the patients in perioperative phase,during postoperative sequential treatment and the period of follow-up between the 2 groups were compared.All data were analyzed by using the covariance analysis,analysis of variance,chi-square test or rank sum test.Results The detection rates of lesions with a diameter less than 1 cm,operation time,liver resection volume,intraoperative blood loss,rate of perioperative blood transfusion,duration of postoperative hospital stay,incidences of postoperative complications were 81％ (22/27),( 186 ± 36) minutes,(75 ± 29) ml,( 189 ± 60) ml,24％(30/126),(21 ± 12)days and 13％ (17/126) in the precise sequential therapy group,and 18％ (2/11),(222 ± 30)minutes,(133 ±88)ml,(327 ±46)ml,51％ (41/81),(26 ± 17)days and 20％ (16/81) in the conventional therapy group,respectively,with significant differences between the 2 groups (F =10.876,7.390,46.996,31.025,14.556,6.315,4.017,P ＜ 0.05).No significant difference was observed on the levels of alanine transaminase,albumin and the Child-Pugh score before and after the intervention in the precise sequential therapy group,but significant differences were observed in the conventional therapy group.The 1-,2-,3-year tumor recurrence rates and the 1-,2-,3-year survival rates were 17％ (21/126),22％ (17/76),26％ (8/31) and 87％ (110/126),87％ (66/76),84％ (26/31) in the precise sequential therapy group,and 31％ (25/81),38％ (27/71),48％(31/65 ) and 77％ (62/81),75％ (53/71 ) and 60％ (39/65) in the conventional therapy group,respectively.There were significant differences in the prognosis of the patients in the 2 groups ( x2 =4.958,4.292,4.168,4.062,3.640,5.470,P ＜ 0.05 ).Conclusion Through accurate assessment of the patients' condition before surgery,precise hepatectomy and precise postoperative intervention,the goal of effective control of tumor recurrence,maximum protection of the liver function and improvement of the survival rate can be achieved.

BACKGROUND:There are most single-center studies about bone marrow stem cels applied to treat decompensated cirrhosis, but the therapeutic results are not ideal. It is possibly related to aging, physical weakness, poor bone marrow hematopoietic function, less available number of stem cels and feeble ability of regeneration and proliferation in liver cirrhosis patients. Umbilical cord mesenchymal stem cels are characterized of easy to obtain, wide source and weak immunogenicity. Co-transplantation of bone marrow stem cels and umbilical cord mesenchymal stem cels may improve the therapeutic effects on decompensated cirrhosis patients. OBJECTIVE:To investigate the efficacy and safety of co-transplantation of umbilical cord mesenchymal stem cels and bone marrow stem cels on decompensated cirrhosis.METHODS:Thirty-two decompensated cirrhosis patients were randomly divided into two groups: in stem cel group, 13 patients received co-transplantation of umbilical cord mesenchymal stem cels and bone marrow stem cels based on regular medical treatment; in control group, 19 patients only underwent the regular medical treatment. Al the patients were folow-up for 1 year. Alanine aminotransferase, albumin, total bilirubin, prothrombin time, Child-Pugh score and Model for End-Stage Liver Disease score, 1-year survival rate, Quality of Life score and adverse reactions related to stem cel therapy were observed and recorded in the two groups at 4, 12, 52 weeks after treatment. RESULTS AND CONCLUSION:At 4, 12, 52 weeks after treatment, improvements in the liver function, prothrombin time, Child-Pugh score and Model for End-Stage Liver Disease score were found in the two groups, but there was no difference between the two groups (P > 0.05). At 4 weeks after transplantation, the clinical symptoms and Quality of Life score in the stem cel group were significantly improved, which were better than those in the control group (P < 0.05). But at 12 and 52 weeks after treatment, no difference was found between the two groups (P > 0.05). In addition, the 1-year survival rate showed no difference between the two groups, and no severe adverse reactions related to stem cel therapy occurred during the folow-up. Co-transplantation of umbilical cord mesenchymal stem cels and bone marrow stem cels is safe and effective to improve the clinical symptoms of decompensated cirrhosis patients. However, further studies with larger samples are warranted to better clarify the co-transplantation effects.

Objective To study comprehensive nursing meassures for small area of diabetic foot ulcer. Methods A total of 58 patients with small area of diabetic foot ulcer in our department were divided into two groups.The comprehensive nursing group (CN group,30 cases) adopted comprehensive nursing and treatment as follows, surgical dcbridement, infrared radiation of wound surface,sprinkling of growth factors(bFGF and rhEGF),covered by hydrocolloid dressing(Algoplaque).The control group(28cases) received hydropathic compress with 0.5% povidone iodine or 5% furacillin solution.Wound heal-ing progreasion,average healing time and the positive rate of wound germiculture of the two groups were observed. Results The average healing time of the CN group was (29.6±9.4)d,which was shorter than that of the control group,( 39.2±13.5 )d, P < 0.05. Statistical difference existed in the positive rate of wound germiculture at the 7th day and the 14th day between the two groups (P < 0.05). Conclusions Routine medical treatment and general nursing measures plus surgical debridement,infrared radiation of wound sur-face,sprinkling of growth factors and hydrocolloid dressing possess facilitative healing effect for small area of diabetic foot ulcer.

Objective To analyze the risk factors of acute kidney injury after orthotropic liver transplantation for adult benign end-stage liver disease.Methods A retrospective analysis was conducted in 30 recipients (18 males,12 females,aged 23-68 years,ASA grade Ⅲ or Ⅳ) who underwent orthotropic liver transplantation for benign end-stage liver disease at Tongji Hospital from May,2014 to December,2014.Both demographic characteristics and perioperative parameters were collected,including general condition,surgery and anesthesia factors and intraoperative salvage autotransfusion or not.Perioperative laboratory findings related to renal function including urine volume,serum creatinine (Scr) and urea nitrogen (BUN) were collected,too.All variables tested in the univariate analysis with a P<0.10 were included in a multiple logistic regression analysis.Results There were less intraoperative salvage autotransfusion,more platelet transfusion and a higher using rate of vasopressors in the AKI group after surgery than those did not.Patients who received intraoperative salvage autotransfusion had 0.058 time odds (95%CI 0.005-0.649) of AKI than those did not;patients who required platelet transfusion had 10.706 times higher odds (95%CI 1.212-94.963) of AKI than those did not.Conclusion It is likely that intraoperative salvage autotransfusion was able to decrease the morbidity of AKI,while platelet transfusion and vasopressor administration to maintain blood pressure could increase the possibility of AKI.