Recent years,mass spectrometer has been extensively used in many clinical laboratories with the development of mass spectrometry technique.Liver diseases are common digestive system diseases and it is harmful to humans' health.Mass spectrometry technique has already been used in identification and genotypes of hepatitis virus and discovery of biomarkers of chronic liver disease such as liver cirrhosis and hepatocellular carcinoma.In this paper,the applications of mass spectrometry in the diagnosis of liver disease were reviewed.

Primary hepatic carcinoma is one of the most common cancer in China.There is no accurate and reliable serological diagnostic method could be used for the early diagnosis and prognosis evaluation of primary hepatic carcinoma at present.The detection of autoantibody for tumor associated antigens has high sensitivity and specificity to diagnose cancer.Thus,measurement of autoantibodies of primary hepatic carcinoma is expected to achieve early diagnosis and prognosis evaluation.The research situation and application prospect of these biomarkers are summarized.

Hepatocellular carcinoma (HCC) is a common and highly lethal cancer which has high morbidity in Asia-Pacific region,especially in china.Early diagnosis is helpful to treatment and prognosis of HCC.Based on the research development of serum tumor markers of HCC in recent years,several promising candidate diagnostic markers were listed and the application of some new technologies was discussed in this field.

The presence of many autoantibodies in the blood of asymptotic patients for several years before the onset of autoimmune diseases,particularly in those who are at high genetic risks.Many of the autoantibodies have high positive predictive value on disease development,disease severity and organ damage,respectively.

Objective To investigate the intracellular and extracellular affinity between hepatitis B virus e antigen (HBeAg) and protein AK026018. Methods The gene AK026018 was amplified by reverse transcription polymerase chain reaction (RT-PCR) technique from HepG2 cell. The expressive vector of pGADT7-AK was constructed by routine molecular biological methods. The auxotroph yeast cells were cotransfected with pGADT7-AK and pGBKT7-eAg and plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-?-gal. To further prove the affinity between HBeAg and protein AK026018, translation was performed by using reticulocyte lysate, and immunoprecipitation was showed in vitro. Results The expressive vector was constructed and confirmed by DNA sequencing analysis and restriction enzyme digestion.The positive clones could grow on SD/-Trp-Leu-His-Ade/x-?-gal medium and were blue. The radioautographic band were found in immunoprecipitation analysis. Conclusions HBeAg could bind protein AK026018 in vivo and in vitro.

Objective To construct prokaryotic expression vector carrying Japanese encephalitis virus (JEV) NS1 gene, and to express the vector in E. coli, so to lay a foundation for the further development of JEV early diagnosis kit. Methods The NS1 gene was amplified by RT-PCR, the target gene and prokaryotic expression plasmid pET28a(+) were digested by BamH I and Hind Ⅲ respectively. The target gene was then purified by DNA extraction kit. A 1∶3 molar ratio of vector: insert DNA ligated with T4 DNA ligase to construct the recombinant plasmid pET28a(+)-NS1. The ligated products were transformed into E. coli BL21 (DE3) and the colonies were selected on LB medium with karamycin. After cultivation, positive colonies were picked out and the recombinant plasmid were identified by endonuclease digestions, PCR rand sequencing. The target protein was expressed with induction of IPTG. The expressed proteins mentioned above were then identified and analyzed by SDS-PAGE and Western-blotting respectively. Results The sequencing results of amplified products showed that JEV-NS1 RNA fragments were about 1 300bp in length which were similar as respected. Compared with the published sequence of SA14-14-2 with Blast, the homology of the nucleotide sequence was 100%. The molecular weight of expressed protein was about 45kD, the result of Western blotting proved the specific antigenicity of the protein. Conclusion The specific JEV nonstructural protein 1 is expressed in E. coli successfully and shows high specificity to the antibody. The stable expression of the protein and the analysis of its antigenic specificity provide foundation to develop the early stage diagnosis kit.

Point-of-care testing ( POCT ) is a new area in the laboratory medicine.This article discusses the current situation in regulations , industry management , quality management , personnel management and training and other aspects of POCT in America and China.Then this article points out the challenges of POCT development in China , and further puts forward advanced measures and suggestions for improving POCT management system in China.

Next generation sequencing(NGS)has made great progress, applied from science technology to clinical diagnosis in the past several years.With a simple, fast, high-throughput, and high-resolution feature, it has been believed to be one of the most potential technology in clinical tests, especially in individual tumor diagnosis and treatment, noninvasive prenatal screening, genetic disorders screening field.This paper will briefly review the application of next generation sequencing in the field of infection disease.

ObjectiveWE transfected the recombinant expression plasmid of pcDNA3. 1-HIF-1α into the prostate cancer cells, to research the effect of HIF-1α on proliferation of prostate cancer cell PC-3.MethodsWe selected a stable expression cell line with G418 were selected by transfection of the recombinant expression plasmid of pcDNA3. 1-HIF-1α into the prostate cancer. The protein and mRNA expression of HIF-1α was assayed by western - blot and RT-PCR. The cells growth curves were described by MTT and the ability of invasion was assayed by Transwell.ResultsThe expression of HIF-1α mRNA was not obviously increased compared to the untransfected prostate cancer cell by RT-PCR, but the expression of HIF-1α protein was up-regulated by western-blot after the recombinant expression plasmid transfected into PC-3. The ability of cell proliferation and invasion was significantly enhanced by MTT and Transwell assays.ConclusionThe stable expression cell model of HIF-1α was successfully constructed, which enhanced the proliferation and invasion of prostate cancer cell PC-3.

Objective To explore the content change of Alpha Fetal Protein(AFP) and Alpha Fetal Protein Variants(AFP-L3) pre-and post-liver transplantation operation for liver cancer patients.Methods AFP-L3 was de-termined by micro centrifugal column from twelve cases of successive sera(For each case,one original shale pre-oper-ation,and mole shares post-operation successively),and preserved from patients who were diagnosed as fiver cancer and completed fiver transplantation operation.simultaneously the content of AFP and AFP-L3 in the original serum were also determined.and the ratio of AFP-L3 in AFP was calculated in order to compare the content change of AFP and the ratio of AFP-L3 pre-and post-liver transplantation operation.Results Compared with twelve cases of pre-and post-operation,the AFP in only five cases turned tO negative;The AFP in other cases keeped positive though AFP-L3%tumde to negative.and the content of AFP in some specific cases still keeped in a high level.Regarding AFP-L3,besides two casses turning to negative immediately after operation,the content of AFP-L3 in other patients de-creased gradually after operation,and it approached to negative(AFP-L3%<10%).Through statistical analysis,be-tween the half lives of AFP and AFP-L3%,there waa no statistic significance.Therefore,AFP-L3% could be considered as an independent reference item.Conclusion After a radical cure of liver resection or liver transplantation operation,AFP-L3%usually turned to negative in a rather short period (about 1 to 20 days).The ratio of AFP-L3 in AFP became be-low 10%,and it indicated that the operation would be quite complete,remaining liver would have hepatitis or hepatocirrho-sis.Otherwise,the ratio was not decrease during a long period,and it indicated the operation had not eliminated focuses completdy.