Objective:To develop an inducible system for expression of GraB in E.coli for the purpose of investion of its function on genome DNA fragment.Methods:Amplified granzyme B cDNA by RT-PCR via extracting lymphocyte total RNA.Identified with endonuclease cut and DNA sequence analysis,the gene was inserted into E.coli expression vector pTrcHis2A,the pokaryotic expression plasmid pTrcHis2A/GraB was constructed and transformed into TOP10F.Results:After 8 h of IPTG in duction, the GraB was expressed 15% of total proteins by SDS-PAGE.Western blot assay proved the expressed hGraB to be of good antigenicity and high specificity.The recombinant protein purified by affinity chromatography has effect on GraB substrate.GraB was found to work directly on the genome DNA fragments,promoting its split.Conclusion:It is found that there may exist certain novel pathway for GraB-induced apoptosis.

Objective To observe the changes of image quality after adjusting the space location and the scan parameters of MRS.Methods 65 patients were scanned with magnetism resonance spectrum head examination.The location,size and other scan parameters of the region interested were changed intentionally.The quality of the images was preliminarily evaluated and the relationship between the parameters and the quality of MRS were probed.Results In the 65 cases,60 were with better shape of wave lines and metabolite peak positions,5 with aberrant lines and shifted peaks.Among the recorded data: the range of half high width value is 5~23,water suppression level 92~99.With the 60-case group,all those half high width values were within the given range of the system,on the contrast,within the 5-case group half high width values were 7,9,16 and 23,and water suppression levels were 95,96,97.Conclusion High-quality wave spectrum depends on high uniformity of magnetic field.The value of half high width is an important factor that reflects the uniformity of magnetic field.

OBJECTIVETo observe the effect of human vascular endothelial growth factor (VEGF) fragment (3 approximately 4 exon) in E. coli on anti-angiogenesis.

METHODSThrough RT-PCR amplification, endonuclease cut and DNA sequence analysis identification, hVEGF fragment cDNA was inserted into E. coli expression vector pTrcHis2A. The prokaryotic expression plasmid pTrcHis2A/VEGF(3 approximately 4) was constructed and transformed into TOP10F.

RESULTSAfter 8hr isopropy-beta-D-thiogalactoside (IPTG) induction, VEGF fragment was expressed in 15% of total proteins through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The expressed protein was highly antigenic and specific. The VEGF fragment was further purified by affinity, which could inhibit HUVEC proliferation and neovascularization of the chick chorioallantoic membrane.

CONCLUSIONVEGF fragment is anti-angiogenetic, which may potentially be used in oncologico-biological targeting therapy.

Angiogenesis Inhibitors ; isolation & purification ; pharmacology ; Cloning, Molecular ; Endothelial Growth Factors ; genetics ; isolation & purification ; pharmacology ; Escherichia coli ; genetics ; Gene Expression ; drug effects ; Genetic Vectors ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; isolation & purification ; pharmacology ; Isopropyl Thiogalactoside ; pharmacology ; Lymphokines ; genetics ; isolation & purification ; pharmacology ; Peptide Fragments ; genetics ; isolation & purification ; pharmacology ; Plasmids ; genetics ; Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

Objective To analyze the risk factors of postoperative recurrence and metastasis of of BCLC stage A (BCLC-A) hepatocellular carcinoma (HCC).Methods Data of 210 cases of liver resection for HCC were retrospectively analyzed from Nov 2013 to June 2016.Multivariate logistic regression analysis was used to analyze the risk factors for postoperative recurrence.The cumulative survival rates were calculated using the Kaplan-Meier method and the diffrences of the related factors between various groups were analyzed by Log-rank test.Results Univariate analysis showed that alpha-fetoprotein (AFP),neutrophil-lymphocyte ratio (NLR),hepatitis B virus surface antigen (HBsAg) positive and intraoperative transfusion were associated with early recurrence and metastasis after hepatectomy (P ＜ 0.05).Multivariate analysis showed that AFP ＞ 400 ng/ml is an independent risk factor for early postoperative recurrence and metastasis (P =0.008).HBV DNA positive can affect the disease-free survival rate after HCC resection (P =0.030).Conclusion AFP is an independent risk factor for early postoperative recurrence and metastasis.Preoperative HBV DNA positive significantly decreases the disease-free survival rate in HCC patients.

Objective To study the value of serum prealbumin-bilirubin score (PALBI) in predicting posthepatectomy liver failure (PHLF) for patients with HBV related hepatocellular carcinoma (HCC).Methods A retrospective study was conducted on 919 HBV-related HCC patients who underwent hepatectomy from September 2013 to December 2016 at the Affiliated Tumor Hospital of Guangxi Medical University.These patients were divided into a training cohort (n =689) and a validation cohort (n =230) using the 3 ∶ 1 matching principle.The training cohort was divided into the control group (n=546) and the PHLF group (n=143) according to whether PHLF occurred.The multivariate logistic regression model was used to analyze the factors related to PHLF in the training cohort,and then the PALBI score was established.The ability of the PALBI score to predict PHLF was evaluated by the area under the receiver operating characteristic curve (AUC) and compared with the Child-Pugh,model for end-stage liver disease (MELD),and albumin-bilirubin (ALBI) scores.Results Univariate and multivariate logistic regression analyses showed the factors including HBV-DNA≥ 103 IU/ml,total bilirubin,prealbumin,platelet count,AST,prothrombin time,intraoperative blood loss ≥400 ml and major liver resection were closely related to PHLF.The ability of the PALBI score (AUC =0.733) to predict PHLF preoperatively was superior to the ChildPugh score (AUC =0.562),the MELD score (AUC =0.652) and the ALBI score (AUC =0.683) in the entire training cohort.Similar results were obtained in the entire validation cohort (AUC:0.752 vs.0.599 vs.0.641 vs.0.678).To eliminate the effect of a small residual liver volume on PHLF,the ability of each of these scores in the training and validation cohorts to predict PHLF was calculated respectively in these 2 cohorts of patients who underwent only minor liver resection,and similar results were obtained.Conclusion The PALBI score was significantly superior to the Child-Pugh,MELD and ALBI scores in predicting PHLF in patients with HBV-related HCC who underwent liver resection.The PALBI score is a simple,non-invasive and reliable novel model in predicting PHLF.