Long-Term Persistence of IgG Antibodies in SARS-CoV Infected Healthcare Workers

BACKGROUNDThe ongoing worldwide outbreak of the 2019-nCoV is markedly similar to the severe acute respiratory syndrome (SARS) outbreak 17 years ago. During the 2002-2003 SARS outbreak, healthcare workers formed a special population of patients. Although virus-specific IgG play important roles in virus neutralization and prevention against future infection, limited information is available regarding the long term persistence of IgG after infection with SARS-like coronavirus. METHODSA long-term prospective cohort study followed 34 SARS-CoV-infected healthcare workers from a hospital with clustered infected cases during the 2002-2003 SARS outbreak in Guangzhou, China, with a 13-year follow-up. Serum samples were collected annually from 2003-2015. Twenty SARS-CoV-infected and 40 non-infected healthcare workers were enrolled in 2015, and their serum samples were collected. All sera were tested for IgG antibodies with ELISA using whole virus and a recombinant nucleocapsid protein of SARS-CoV, as a diagnostic antigen. RESULTSAnti SARS-CoV IgG was found to persist for up to 12 years. IgG titers typically peaked in 2004, declining rapidly from 2004-2006, and then continued to decline at a slower rate. IgG titers in SARS-CoV-infected healthcare workers remained at a significantly high level until 2015. Patients treated with corticosteroids at the time of infection were found to have lower IgG titers than those without. CONCLUSIONSIgG antibodies against SARS-CoV can persist for at least 12 years. The presence of SARS-CoV IgG might provide protection against SARS-CoV and other betacoronavirus. This study provides valuable information regarding humoral immune responses against SARS-CoV and the 2019-nCoV.

Analysis of Differentially Expressed Proteins Involved in Autoimmune Cirrhosis and Normal Serum by iTRAQ Proteomics.

PURPOSE: In order to study the candidate biomarkers in autoimmune cirrhosis (AIC). EXPERIMENTAL DESIGN: Isobaric tags are first implemented for relative and absolute quantitation technology on proteins prepared from serum obtained from AIC and normal controls. Proteins found to be differentially expressed are identified with liquid chromatography electrospray ionization tandem mass spectrometry by using a Q Exactive classic ion trap mass spectrometer. RESULTS: 108 proteins (32 upregulated and 76 downregulated proteins) are identified from AIC samples, compared with the normal controls. Gene Ontology enrichment analysis, KEGG pathway analysis, and protein-protein interaction map by STRING show that they associate with multiple functional groups, including ion binding activity, peptidase activity, and enzyme regulator activity. Finally, the von Willebrand factor, insulin-like growth factor-binding protein complex acid labile subunit, transthyretin, adiponectin proteins are identified with western blot as candidate biomarkers for AIC. CONCLUSIONS AND CLINICAL RELEVANCE: These findings offer a comprehensive profile of the AIC proteome about candidate biomarkers and provide a useful basis for further analysis of the pathogenic mechanism of AIC.

A family of Alport syndrome with a novel dominant hereditary hematuria and a mutation in the COL4A4 gene / 中华检验医学杂志

Objective Alport syndrome was an inherited kidney disease caused by the mutation of COL4A3,COL4A4, or COL4A5. Whole-exome sequencing was used to detect the mutations on these genes for the molecular diagnosis of Alport syndrome. Methods A 6-year-old girl found accidentally with microscopic hematuria at the age of 4. The clinical data and blood sample of the family including proband, parents, brothers, and sisters were collected. Whole exome sequencing was conducted using their genomic DNAs. Results A novel heterozygous frameshift mutation c.1826delC (p.Pro609Glnfs*44) was found in the exon 25 of the COL4A4(NM_000092) in the proband, the father, and the sister, showing an autosomal dominant inheritance pattern of Alport syndrome. This mutation of COL4A4 was confirmed by mutation analysis, and the mutation of c.1826delC was verified by Sanger sequencing. No mutations on COL4A3 and COL4A5 were detected in this family. And the mother and brother are normal wide-type. Conclusions This novel mutation is a valuable addition to the current genetic profile of Alport syndrome, and provide us a better understanding of the disease. Whole-exome sequencing is a power tool to identify the novel mutations of inherited disease and contribute to the molecular diagnosis of disease.

The development and prospect of the detection technology of circulating tumor cells and circulating tumor DNA / 中华检验医学杂志

Circulating tumor cells (CTC) and circulating tumor DNA(ctDNA)are cancer cells that detach from their primary site to circularity system or other body fluids,which hold the promise of monitoring of tumor evolution and therapeutic efficacy and improving prognosis for patients.With the rapid development of detection technologies of CTCs and ctDNA,the sensitivity and specificity of current assays had significant improved.CTCs can be detected by use of cytometry,immunofluorescence analysis and genedetection and the detection methods of ctDNA mainly based on PCR and the next generation sequencing.As the developing of various methods and platforms were developed for detecting the circulating tumor cells and circulating tumor DNA,it is necessary to use the detection technology flexibly according to the research purpose and provide more assistance for the diagnosis and treatment of tumor.

A family of Alport syndrome with a novel dominant hereditary hematuria and a mutation in the COL4A4 gene / 中华检验医学杂志

Objective@#Alport syndrome was an inherited kidney disease caused by the mutation of COL4A3, COL4A4, or COL4A5. Whole-exome sequencing was used to detect the mutations on these genes for the molecular diagnosis of Alport syndrome.@*Methods@#A 6-year-old girl found accidentally with microscopic hematuria at the age of 4. The clinical data and blood sample of the family including proband, parents, brothers, and sisters were collected. Whole exome sequencing was conducted using their genomic DNAs.@*Results@#A novel heterozygous frameshift mutation c.1826delC (p.Pro609Glnfs*44) was found in the exon 25 of the COL4A4 (NM_000092) in the proband, the father, and the sister, showing an autosomal dominant inheritance pattern of Alport syndrome. This mutation of COL4A4 was confirmed by mutation analysis, and the mutation of c.1826delC was verified by Sanger sequencing. No mutations on COL4A3 and COL4A5 were detected in this family. And the mother and brother are normal wide-type.@*Conclusions@#This novel mutation is a valuable addition to the current genetic profile of Alport syndrome, and provide us a better understanding of the disease. Whole-exome sequencing is a power tool to identify the novel mutations of inherited disease and contribute to the molecular diagnosis of disease.

Application of SAT technique in detection of Chlamydia trachomatis and Ureaplasma urealyticum / 中华检验医学杂志

Objective To study the Chlamydia trachomatis(CT)and Ureaplasma urealyticum (UU)infection in Guangzhou area, and analyze the consistency of simultaneous amplification and testing (SAT)and conventional methods(CT was detected by latex immunochromatography, UU was detected by liquid culture method).Methods A total of 12 120 samples of urogenital secretions or urine samples were collected from Sun Yat-Sen Memorial Hospital of Sun Yat-Sen University from January 2015 to December 2016.CT-RNA and UU-RNA were detected by the SAT technique, a part of samples were tested by conventional methods at the same time.The positive rates of CT and UU by SAT and the conventional methods between different gender and age groups were analyzed by χ2test, the consistencies between different detection methods were analyzed by Kappa test.Results The positive rate of CT was 4.05%(356/8 781), UU 33.69%(1 125/3 339)in Guangzhou from 2015 to 2016.The positive rate of UU was significantly higher than that of CT(χ2=1 981,P<0.01).Of 145 specimens for CT test,the coincidence rate between SAT and latex immunochromatographic method was 96.55%(140/145), which showed good consistency(Kappa=0.65).Of 186 specimens for UU test,the coincidence rate of the results between the SAT method and liquid culture was 92.47%(172/186),which showed strong consistency(Kappa=0.81). Conclusions The positive rate of UU was significantly higher than that of CT in Guangzhou.The SAT method and conventional methods to detect CT and UU show high consistency, which can provide the evidence for clinical diagnosis of CT and UU infection.

Establishment of the test method of the Aβ42 in the serum of the alzheimer disease patient and its clinical significance / 中华检验医学杂志

ObjectiveTo establish a enzyme-linked immunosorbent assay (ELISA) method for detecting the β-amyloid peptide 42 ( Aβ42 ) and explore its clinical meaning for diagnosis and treatment in the early stages of the alzheimer disease ( AD).Methods Using the Aβ42 single chain variable fragment constructed by phage antibody library display system as coat antibody,associated with the Aβ42 polyclonal antibody acquired by Aβ42 immunized rabbit and HRP labeled goat anti rabbit IgG to establish ELISA method for detecting the Aβ42 in peripheral blood.The method was used it to test the Aβ42 in 120 vascular dementia VD) or cerebral vessel infarction patients and 120 AD patients and 120 controls.The methodology performance were evaluated.ResultsThe inter and intra coefficient of variable (CV) of this self-established ELISA method was 3.6％ and 3.5％,6.8％ and 7.1％ respectively.The recovery rate was 97.2％ -103.1％.The linear range was 0.050 - 2 μg,/L.Its reactivity decreased ＜ 12％ when it was put in both 37 ℃ for 6 days and 4 ℃ for 6 months.Compared with the Belgium INNOTEST reagent by testing 90 samples simultaneously,the results of self-established method was (0.207 ± 0.039 ) μg/L,the results of INNOTEST was (0.206± 0.038 ) μg/L; the regression equation was Y =1.011X - 0.003,R2 =0.979,P ＜0.01.The Aβ42 in blood of AD group was (0.247 ± 0.032 ) μg/L,VD or cerebral vessel infarction group was (0.173 ±0.028) μg/L,control group was (0.172 ±0.032) μg/L.The Aβ42 in AD group was higher than that in the VD or cerebral vessel infarction group and control group (q =18.867,18.907respectively,P ＜ 0.01 ).The cut off value was 0.212 μg/L decided by the receiver operating characteristic (ROC) curve.The reference interval was 0 -0.212 μg/L.The sensitivity of this ELISA method was 86.7％(104/120) and specificity was 90.8％ (218/240).ConclusionsThe ELISA method for detecting Aβ42 in peripheral blood established by the study is sensitive and specific and has good precision and stability.It could provide a new effective criterion and support for the early diagnosis and treatment of the AD patients.

WT inhibit human hepatocellular carcinoma BEL-7402 cells growth by modulating Akt and ERK1/2 phosphorylation / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate effects of Akt and ERK1/2 signaling pathways on waltonitone (WT) induced cell growth inhibition in human hepatocellular carcinoma BEL-7402 cells.</p><p><b>METHOD</b>Cell viability of BEL-7402 cells was examined using MTT assay. Phosphorylation of E Akt and RK1/2 were detected by Western blot analysis, while cell cycle distribution of BEL7402 cells was analyzed by flow cytometry.</p><p><b>RESULT</b>WT could inhibit the BEL-7402 cells growth, induce the S-phase cell cycle arrest, activate Akt and ERK1/2 phosporylation. Moreover, the cell growth inhibition and S-phase cell cycle arrest induction of WT on BEL-7402 cells could be blocked by Akt and ERK1/2 inhibitors.</p><p><b>CONCLUSION</b>WT induce the cell cycle arrest and inhibit the cell growth on BEL-7402 cells by modulating Akt and ERK1/2 phosphorylation.</p>