With the improvement of the multi-parameter and high throughput technology,FCM has expanded its clinical application from cellular phenotying to other areas,such as protein function analysis,fine needle aspiration,cerebrospinal fluid determination,circulating tumor cell detection as well as clinical microbiology.Currently,lack of standards for the sample processing,equipment adjustment and data analysis remains the main obstacle for routine application of FCM.Therefore,it is recommended to establish consensus standard operation procedures as well as shared FCM platform regionally to provide better service of FCM in clinical laboratory.

During the past decade, tremendous progress has been made in terms of speed, read length, and throughput, along with a sharp reduction in per-base cost.Together, these advances democratized next generation sequence (NGS) and paved the way for the development of a large number of novel NGS applications in clinical diagnostics, especially in the field of non-invasive prenatal detection, rare genetic disease and cancer companion diagnostics.As technology advances, long-read single molecule sequencing began to emerge.Single cell, long-reads, transcriptome, and low cost will be the NGS direction.Due to the special nature of clinical testing, the current NGS clinical application system,including genetic counseling, testing standards, quality control, supervision, database construction etc, does not match the national conditions well and still faces a few challenges, needs to be constantly improved through the routine clinical practice in the future.

Hepatic fibrosis is the final common pathway for liver injury caused by various chronic liver diseases.Noninvasive assessments of liver fibrosis are valuable in clinical use since they are easy to apply and monitor disease progression.Serologic biomarkers for liver fibrosis consist of direct markers which reflect ECM turnover and indirect markers which show liver function of synthesis,metabolism and reservation.A number of assessment models can identify significant fibrosis and cirrhosis,and avoid unnecessary liver biopsy.They can be used for disease staging,prognosis prediction,and treatment surveillance.Variation of analytical performances exists among diverse analyzers and assays.The standardization of these tests will enhance their comparability.Large scale and multicenter clinical verification studies are needed for the clinical utility of these assessment models based on Chinese population of chronic hepatic diseases.

Molecular diagnosis is rapidly developed in recent years , mainly applicated in the diagnosis of hereditary disease , infectious pathogens, tumor susceptibility and molecular typing , companion diagnosis and prognosis assessment , playing more and more important role in many diseases diagnosis and treatment.Molecular diagnosis was developed from the eighties of the last century in our country .Nowadays, the mainly applied technologies in the clinical laboratory include fluorescence in situ hybridization , quantitative PCR, microarray and DNA sequencing. These molecular technologies make up for the insufficiency of routine testing and take up a central role in the development of modern laboratory medicine . With the continuous development in transformation research of molecular technology recent years , there will be more molecular diagnostic techniques applied in clinicaldiagnosis in the future .But it still exists some drawbacks in the performance of molecular diagnosis in our country according to the current situation , such as imbalanced regional development , mismatched policies, non-standardized laboratory construction , deficiency of quality control and supervision , etc., which requires the joint effort of the government , hospital, professional association and clinical laboratory itself to promote the healthy and orderly development of molecular diagnosis.

Objective To discuss Foxp3 expression in peripheral blood from the patients with various stages of gastric cancer by SYBR Green I real-time PCR assay.Methods Foxp3 standard was prepared by conventional RT-PCR and employed to generate quantitative PCR standard curve.The precision,sensitivity and specificity of the assay were measured.SYBR Green I real-time PCR assay was adopted to detect and quantify Foxp3 expression in peripheral blood from 40 gastric cancer patients and 8 normal controls.Results The coding sequence (CDS) of full length Foxp3 was obtained and used as standards.The dynamic range of standard curve of real-time PCR assay was 102-108 copies.The mean coefficient of variation (CV) of inter-and intra-assay was less than 5%.The minimum detectable limit was 280 copies of Foxp3 mRNA.Foxp3 expression level in gastric cancer was significantly higher than that in normal control (P

Objective Two Chinese pedigrees with congenital factor Ⅻ(FⅫ)deficiency were enrolled in the present study,and studies on the clinical manifestations,family survey,biochemical examinations and gene diagnosis of these pedigrees were performed.Methods In October 2014-2015 March,two cases of hereditary FⅫ deficiency patients were included in Xinhua hospital.Activated partial thromboplastin time(APTT),FⅫ procoagulant activity(FⅫ:C),FⅫ antigen(FⅫ:Ag)and other parameters of coagulant were detected.The FⅫ deficiency pedigree members,exons 1-14,boundary introns including the splice junctions of the F12 gene were amplified with polymerase chain reaction(PCR).Direct sequencing was exerted to purified PCR product to detect the gene mutation.If the gene mutations were found,polymorphism should be ruled out by directing sequence.One hundred and three healthy persons as normal controls.Results The two probands were manifested prolonged APTT(101 s and 143 s).They showed lower FⅫ activity and FⅫ antigen(2％and 6％,0.4％and 4％,respectively).FⅡ:C,FⅦ:C,FⅧ:C,FⅨ:C,FⅩ:C and Fg are normal in the two probands.LAC is negative.Proband 1 has c.1285C>T(p.Q429 stop)mutation.His parents and son have the heterozygous mutation in the same position.Proband 2 has c.1556T>C(p.L519P)mutation.Her two sons have the heterozygous mutation in the same position.In the promoter regions of F12 gene,there were common 46C/T and 619 G/C polymorphisms in two pedigrees.Conclusion c.1285C>T(p.Q429 stop)and c.1556T>C(p.L519P)are the cause of FⅫ deficiency.

Objective To investigate the clinical value of immunophenotyping and quantitative analysis of serum immunoglobulin in the diagnosis and typing of monoclonal gammaglobulinemia.Methods 118 serum samples were collected from the patients with monoclonal gammaglobulinemia and performed the serum protein electrophoresis(SPE),serum immunofixation(IF)electrophoresis, immunoglobulins(Ig)quantitation and serum light chain κ,λdetection.The analysis was conducted aiming at the patients with im-munophenotyping positive and the Ig quantitation negative.Results 56 cases showed immunophenotyping positive and the immuno-globulin quantitation negative,among them,6 cases were free light chain type (κlight chain in 2 cases andλlight chain in 4 cases), 1 case was the non-secretion type;62 cases showed immunophenotyping positive and the Ig quantitation positive.Conclusion The IF technique has an important significance for the diagnosis and typing of monoclonal gammaglobulinemia,its diagnosis can be com-bined with the other laboratory tests of serum Ig and light chain quantitative detection.

Flow cytometry (FCM) is one of the most advanced cell quantitative analysis technology.Today,flow cytometry has been extensively and intensively used in medicine and other science,ranging from basic research to clinical diagnosis With the rapid development of science and technology of China,the application of flow cytometry in clinical research and diagnosis has also made significant progress.

opping drinking. They were not influenced by gender, smoking and drinking histories. They could serve as monitoring indexes for recent drinking status on healthy individuals.

Objective To explore the levels of serum chemokine in the patients with gastric cancer using cytometric beads array (CBA) and to find out the laboratory evidence for gastric cancer immunotberapy.Methods Forty-five patients with gastric cancer,thirty patients with benign gastric diseases and forty healthy controls were included.The chemokine levels of interleukin-8 (IL-8),regulated upon activation normal T-cell expressed and secreted (RANTES),monokine induced by IFN-γ (MIG),monocyte ehemoattractant protein-1(MCP-1) and interferon-γ induced protein-10 (IP-10) in serum of these three groups were measured using CBA ,then these data were analyzed using BD CBA analysis software.Results The levels of serum chemokines in healthy controls were (176.4±20.7) μg/L for IP-10,(111.3±17.2) μg/L for MCP-1,(503.9±47.2) μg/L for MIG,(472.4±116.7) μg/L for IL-8,respectively.In gastric cancer patients,the levels were (266.6±24.7) μg/L for IP-10,(100.4,70.8-193.5) μg/L for MCP-1,(1614±275.4) μg/L for MIG,(500.0±164.8) μg/L for IL-8.The levels of serum chemokines in patients with benign gastric disease were (207.9±31.7) μg/L for IP-10,(121.2±23.6) μg/L for MCP-1,(514.5±63.0) μg/L for MIG,(480.2±134.8) μg/L for IL-8,respectively.The levels of IP-10 and MIG among these three groups were significantly different (IP-10:F = 3.52,P < 0.05,MIG:F = 9.27,P < 0.01).The levels of IP-10 were elevated in the cancer patients compared with healthy controls (P < 0.05),while the levels of MIG were elevated significantly in the cancer patients compared with healthy controls and benign disease controls(t=3.29,P < 0.01,t=2.84,P<0.01).Conclusions CBA is a novel method with convenience,sensitivity and accuracy for the detection of serum ehemokines.The concentration of IP-10 and MIG in gastric cancer patients are elevated,indicating its implication in cancer pathogenesis and metastasis.Measurement of serum chemokines will lay the groundwork for therapeutic strategy in gastric cancer by inhibiting and anti-chemokine.