A multicenter study to develop and validate a novel C-GALAD Ⅱ HCC prediction model based on serological markers / 中华检验医学杂志

Objective:To establish a model C-GALAD for detecting hepatocellular carcinoma (HCC) from the chronic liver disease and healthy people based on the serum markers.Methods:A clinical cohort including 229 hepatocellular carcinoma patients, 2 317 patients with chronic liver disease and 982 healthy people, was retrospectively collected from eight hospitals or physical examination institutions from April 2018 to October 2020. The data were divided into a training set and a testing set by stratified sampling with a 6∶4 ratio. A predictive model was established on the training set using a logistic backward regression method and validated on the testing set. In addition, clinical data from March to July 2021 in Beijing You′ an Hospital affiliated to Capital Medical University, including 84 patients with liver cancer and 204 patients with chronic liver disease collected were used for external independent validation of the model. The receiver operating characteristic curve (ROC) area under curve (AUC), the sensitivity and the specificity were used to evaluate the effectiveness of the model.Results:Through the logistic backward regression method, the seven signatures including age, gender, alpha-fetoprotein (AFP), alpha-fetoprotein alloplasm-3 ratio (AFP-L3%), des-gamma-carboxyprothrombin(DCP), platelet (PLT) and total bilirubin (TBIL) were selected as risk factors in the detection model. The area under the ROC curve (AUC) of the model on the testing set was 0.954, with an 88.04% sensitivity and a 94.85% specificity, and the AUC of model on the external independent validation set was 0.943, with an 89.29% sensitivity and a 90.2% specificity, which were better than other published models.Conclusion:The C-GALAD Ⅱ model can accurately predict the risk of hepatocellular carcinoma occurrence, and thus provide a trustworthy diagnosis method of hepatocellular carcinoma.

Clinical diagnostic value of autoantibody detection in autoimmune diseases / 中华检验医学杂志

Autoimmune disease is a kind of immune dysfunction disease caused by gene and environmental factors. At present, there are some difficulties in early diagnosis, prognosis evaluation and curative effect evaluation. More and more autoantibodies have been found and confirmed to be closely related to the occurrence and development of autoimmune diseases with the deepening of related research. Accurate autoantibody detection is helpful to improve the diagnostic efficiency, therapeutic effect and quality of life of patients with autoimmune diseases.

Advances in 2019 novel coronavirus research and laboratory testing / 中华检验医学杂志

Studies have confirmed that the outbreak was caused by a new type of coronavirus infection, which is highly contagious. In this paper, the research progress of coronaviruses and 2019 novel coronavirus as well as the laboratory testing and related biosafety requirements of 2019 novel coronavirus are reviewed to provide relevant basis for the work of medical laboratories and testing institutions.

The new demands for HCV related clinical examinations for DAA treatment / 中华检验医学杂志

Hepatitis C virus (HCV) is one of leading causes for chronic liver diseases, the treatment area has been changed rapidly since direct acting antiviral agents (DAAs) were approved to treat patients with chronic HCV infection. To cure hepatitis in 2030 is the goal set by World Health Organization. However, globally, especially in China, there are still many difficulties needing to be resolved on the cure road of chronic hepatitis C, such as large patient population, high percentage of untreated patients, nonstandard DAA treatment, low efficacy of special groups, and resistance-associated variants in baseline, etc. This comment will focus on the new demands for HCV related clinical examinations for DAA treatment, so as to improve the treatment effectiveness and to help achieve the goal of eliminating viral hepatitis .

The clinical application of laboratory tests and diagnosis for common chronic liver diseases / 中华检验医学杂志

Chronic liver disease has a high incidence and prevalence in China, which had done great harm to people's health. Laboratory detection is an effective way to diagnose and evaluate therapeutic effect of chronic liver disease. However, the selection and interpretation of laboratory tests are difficult for the various causes of liver diseases. According to etiology of the chronic liver diseases, application of common laboratory tests for liver diseases based on the clinical guidelines were interpreted in this article .

Current developments of biosensors-based hemoglobin A1e detection and its POCT analyzers in the market / 中华检验医学杂志

Measurement of glycosylated hemoglobin (HbA1c) is an important indicator for assessing long term glycemic control in individuals with diabetes mellitus.There are more than 300 different assay methods for the analysis of HbA1c.This review outlines the current developments about biosensors assay as well as the situation of POCT analyzers in the market for HbA1c detection at present.

Application of next generation sequencing technology in clinical diagnosis of infectious disease / 中华检验医学杂志

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.

Difference and clinical significance of serum GP73 between alcoholic liver disease and non-alcohol fatty liver disease / 中华实验和临床病毒学杂志

Objective@#The present study was designed to evaluate the clinical significance of serum GP73 in patients with alcohol liver disease.@*Methods@#Thirty-two patients with alcoholic liver disease (ALD), 40 patients with non-alcohol fatty liver disease(NFLAD), and 40 apparently healthy individuals were included in this study.@*Results@#Compared with Those of healthy control population(43.91±19.02 ng/ml), the serum GP73 levels in ALD patients(93.39±66.91 ng/ml) were significantly increased, and also markedly higher than those of NFLAD patients (55.38±21.00 ng/ml). Taken the NFALD as"healthy controls" population, and ALD population as"patients" , the GP73 may be used to differentiate the ALD from NFLAD. The area of ROC analysis was 0.66(95%CI: 0.52~0.80, P=0.02). We set 75.65 ng/ml as the cut-off value for ALD diagnosis, the sensitivity and specificity were 50.0% and 85.0% respectively.@*Conclusion@#Serum GP73 levels in ALD patients were significantly higher than those of NFALD patients. GP73 may be a new marker for differentiating ALD from NFALD.

The research progress on diagnostic values and mechanism of complements in liver diseases / 中华检验医学杂志

The complement system is a part of the immune system and many of the serum complement components are synthesized primarily in the liver.Furthermore,the complement system is also involved in the immune mechanism of pathologic process of liver diseases.The determination of serum complement components level has high clinical diagnostic values for liver diseases.It summarizes the research progress of diagnostic values and mechanism of complement system in liver diseases.

The significance of autoantibodies detection for the diagnosis of primary hepatic carcinoma / 中华检验医学杂志

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.

The research development of mass spectrometry technique in diagnosis of liver disease / 中华检验医学杂志

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.

Analysis of application and management status of point-of-care testing in China and America / 中华检验医学杂志

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.

The predictive value of autoantibodies in autoimmune diseases / 中华检验医学杂志

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.

Research advance of serum diagnosis in hepatocellular carcinoma / 中华检验医学杂志

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.

Effect of hypoxia-inducible factor-1α on proliferation and invasion of prostate cancer PC-3 cell in hypoxic situation / 中华泌尿外科杂志

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.

The content change of AFP and AFP-L3 pre-and post-liver transplantation operation for liver cancer patients / 中国基层医药

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.

Isolation and identification of the viral pathogen in an incident with crowd fever of adults / 中华微生物学和免疫学杂志

Objective To isolate and identify the causative agent in an incident with crowd fever of adults in Henan province.Methods The cells was inoculated by the throat swabs of the patients and followed by neutralization assay and other molecular methods.Then indirect immunofluorescence assay was performed to detect the specific IgM/IgG antibodies against virus in the serum of the patients.Results We have isolated 2 strain adenoviruses from 10 swab samples,which were both identified as Ad11 by sequence analysis and neutralization test.6 of 10 samples were positive fer IgM specific for adenovirus and 3 positive for IgG.The remaining sample was negative for both.Conclusion The causative agent in this incident with crowd fever of adults was Ad11 in group B.

Sero-and molecular epidemiology of the gene of different ORF of transfusion transmitted virus (TTV) in different population of China / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To investigate the distribution of anti-TTV antibody and the different ORF gene in different populations of China.</p><p><b>METHODS</b>The antibody to TTV in sera collected from different population were detected by using ELISA and the different ORF genes were amplified with PCR.</p><p><b>RESULTS</b>The positive rates of TTV ORF1 DNA, ORF2 DNA and the antibody in various populations were as follows: 16.0% (12/75), 10.7% (8/75) and 25.3% (19/75) in paid blood donors; 10.0 (3/30), 16.7% (5/30) and 16.7% (5/30) in patients with hepatitis A, 47.5% (19/40), 42.5% (17/40) and 22.5% (9/40) in patients with hepatitis B; 42.9% (15/35), 37.1% (13/35) and 28.6% (10/35) in patients with hepatitis C; 20.0% (3/15), 26.7% (4/15) and 13.3% (2/15)in patients with hepatitis D; 16.7% (2/12),16.7% (2/12) and 33.3% (4/12)in patients with hepatitis E; 23.8% (5/21), 38.1% (8/21) and 23.8% (5/21) in patients with hepatitis G; 61.1% (11/18), 50.0% (9/18) and 44.4% (8/18) in patients with non A-G hepatitis, respectively. The positive rate of different ORF DNA had no significant difference. Significant differences were found in the positive rates of TTV DNA in various populations. There was no relationship between the TTV DNA and the antibody to TTV.</p><p><b>CONCLUSIONS</b>The antibody to TTV and TTV DNA were found in every population of China. There was no significant difference in the positive rates of TTV DNA between ORF1 and ORF2. The positive rate in patients with non A-G hepatitis was higher than those in the other populations.</p>

Affinity between hepatitis B virus e antigen and protein AK026018 / 中华传染病杂志

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.

High expression of the nonstructural 1 genes of Japanese encephalitis virus (JEV) strain SA14-14-2 in E.coli and the antigenicity analysis of its expressed products / 解放军医学杂志

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.