Reference Intervals of Fibrosis Index Based on Four Indicators in Healthy Elderly Chinese.

BACKGROUND: The establishment of reference intervals of total bilirubin, Alanine aminotransferase (ALT), aspartate aminotransferases (AST), and creatinine provided necessary reference in screening and diagnosis of various kidney and liver diseases. However, these reference intervals were not available to estimate liver fibrosis degree. The purpose of this study is to establish the reference intervals of fibrosis index based on the four indicators (FIB-4) in apparently healthy elderly Chinese. METHODS: A total of 24,949 blood specimens were collected by the standard procedures, and ALT, AST, and PLT were determined. FIB-4 were calculated by the following formula: FIB-4 = (age [years] × AST [U/l])/((PLT [10(9) /l]) × (ALT [U/l])(1/2) ).The elderly's FIB-4 were analyzed between the same age of different sexes and different ages of the same sex. Statistical data were analyzed by SPSS18.0 software. RESULTS: Reference intervals of FIB-4 index, established for the healthy elderly, were 0.9923-4.5424 for males and 0.9007-4.1934 for females. CONCLUSION: We established reference intervals of FIB-4 index. This research provided reference value that can be used by relevant clinicians and inspection officers.

Laboratory reference intervals of complete blood count for apparently healthy elderly people in Shuyang, China.

BACKGROUND: Currently, there are no appropriate RIs of blood cells available for the elderly in most clinical laboratories in China. The aim of this study is to establish the RIs of complete blood cell count for apparently healthy elderly people. METHODS: Blood specimens were collected from elderly residents by standard procedures. Complete blood counts were determined by Sysmex XE-2100 analyzer. The RIs and 95% confidence intervals were calculated by the robust method recommended by CLSI C28-A3 guideline. RESULTS: RIs established for healthy elderly include: total WBC 3.63 - 10.3 x 10(9)/L for males and 3.64 - 10.3 x 10(9)/L for females; RBC 3.74 - 5.49 x 10(12)/L for males and 3.74 - 5.53 x 10(12)/L for females; Hb 109 - 167 g/L for males and 109 - 168 g/L for females; HCT 36.0 - 51.8% for males and 35.7 - 51.8% for females; MCV 86.0 - 105 fL for males and 86.2 - 106 fL for females; MCH 26.4 - 33.6 pg for males and 26.4 - 33.8 pg for females; MCHC 293 - 333 g/L for males and 291 - 335 g/L for females; RDW-SD 39.3 - 53.7 fL for males and 39.6 - 54.5 fL for females; RDW-CV 11.7 - 15.1% for males and 11.7 - 15.2% for females; PLT 122 - 355 x 10(9)/L for males and 122 - 350 x 10(9)/L for females; PCT 14.1 - 37.6 x 10(-1) mL/L for males and 13.9 - 37.9 x 10(-1) mL/L for females; MPV 11.3 - 15.5 fL for males and 11.3 - 15.5 fL for females; PDW 9.74 - 17.0% for males and 9.72 - 17.0% for females; platelet-LCR (P-LCR) 21.3 - 51.2% for males and 21.1 - 51.4% for females. CONCLUSIONS: We established scientific and reasonable RIs of blood cell analysis for the healthy elderly in our region.

Reference intervals for total bilirubin, ALT, AST and creatinine in healthy Chinese elderly.

BACKGROUND: The aim of this study was to establish the reference intervals (RIs) of total bilirubin (TBIL), alanine aminotransferase (ALT), aspartate transaminase (AST), and creatinine (CREA) for apparently healthy elderly (Han ethnicity) in Shuyang, China. MATERIAL AND METHODS: A total of 54 912 blood specimens from elderly residents age 65-104 years were collected by standard procedures in Shuyang county of Jiangsu province. TBIL, ALT, AST, and CREA for each participant were determined by automatic biochemical analyzer. Distribution and differences of TBIL, ALT, AST, and CREA were analyzed and compared between the elderly of the same age of different sexes and different ages of the same sex. RIs of TBIL, ALT, AST, and CREA were compared with the current RIs. The RIs and 95% confidence intervals were calculated using nonparametric method (2.5th-97.5th percentiles) according to the guideline of the Clinical and Laboratory Standards Institute. RESULTS: RIs established for the healthy elderly include: TBIL 7.8~30.6 µmol/L for males and 7.3~26.1 µmol/L for females; ALT 8.7~47.3 U/L for males and 8.4~45.2 U/L for females; AST 15.7~46.9 U/L for males and 15.1~46.2 U/L for females; and CREA 45.1~100.9 µmol/L for males and 38.7~85.0 µmol/L for females. Reference intervals of TBIL, ALT, AST, and CREA for male elderly were higher than those of females, and values of CREA increased with increasing age. CONCLUSIONS: We have established a panel of locally relevant RIs. It is necessary to establish scientific and reasonable RIs of TBIL, ALT, AST, and CREA for the healthy elderly in our region, which will provide a reference for clinicians and inspection officers.

[Expression and antigen binding activity of anti-human CD86 diabody].

AIM: To construct a recombinant eukaryotic expression vector of anti-human CD86 diabody gene and express anti-CD86 diabody through Chinese hamster ovary (CHO) cells, then analyze the capability of the diabody to recognize the tumor cells expressing CD86 and its biological effect. METHODS: The antibody heavy and light chain viable region gene (V(H); and V(L);) were cloned from hybridoma cell 1D1 which secreted anti-human CD86 monoclonal antibody. Anti-human CD86 diabody gene V(H);-(GGGGS)-V(L); was constructed by SOE-PCR. Then we inserted it into eukaryotic expression vector to construct a recombinant vector pIRES2-EGFP/CD86-diabody. The recombinant vector was transfected into CHO cells with Lipofectamine(TM); 2000, and the cell clones secreting CD86 diabody were screened by G418. We used IMAC to purify CD86 diabody and quantified its concentration by BCA method. The capability of the diabody to recognize the CD86 expressed on Raji and Daudi was analyzed through flow cytometry. After Raji cells were treated with CD86 diabody for 72 h, its proliferation inhibiting effect was investigated by MTT assay. RESULTS: We have obtained one CHO cell line that stably secreted CD86 diabody. The concentration of CD86 diabody after purification was 5.24 mg/L. The positive rates of CD86 diabody to recognize Raji and Daudi were 77.2% and 70.6%, respectively. After CD86 diabody treatment for 72 h, the inhibition rate of Raji cells was 37%. CONCLUSION: The anti-human CD86 diabody which we obtained successfully could recognize CD86 expressed on tumor cells specifically, and inhibit the proliferation of these tumor cells effectively.

[Effect of CD80-scFv on recognition of CD80 molecule and proliferation of different tumor cells in vitro].

AIM: To prepare CD80-scFv in CHO cells and investigate its effect on the recognition and proliferation of different tumor cells. METHODS: The CD80-scFv was purified from culture supernatant without fetal calf serum (FCS) by IMAC affinity chromatography, and then bound to CD80 molecule on the Daudi, U251, A375 and 8266 cells detected by flow cytometry. Different concentrations of CD80-scFv were added in the training system of different tumor cells, and we analyzed the influence on the proliferation of these cells by MTT assay. With 8266 cells which expressed CD80 highly and were treated by mitomycin firstly as the APC, and human PBMC as the effect cells, we analyzed the influence of the CD80-scFv on the proliferation of PBMC by blocking the co-stimulatory signal. RESULTS: The concentration of CD80-scFv purified from FCS was about 6.67 mg/L and the binding rate of CD80-scFv with Daudi, U251, A375, 8266 and U266 were 96.8%, 39.6%, 20.5%, 99.9% and 3.8%, respectively. CD80-scFv suppressed the proliferation of Daudi and 8266 cells which expressed CD80 highly, and the inhibition rate of Daudi and 8266 cells cultured with CD80-scFv (the final concentration was 25 µg/mL) was 24.04% and 24.16%, respectively. Additionally, the antibody suppressed the proliferation of PBMC by blocking the co-stimulatory signal mediated by CD80-CD28. CONCLUSION: CD80-scFv can recognize CD80 molecule and suppress the proliferation of tumor cells which express CD80 highly.

[Changes in the expression of T lymphocyte subsets in ageing mice induced by D-galactose].

AIM: To explore the changes and significance of spleen T cell subsets in ageing mice induced by D-galactose. METHODS: Ageing mice model was successfully established by 100 g/L D-galactose. The content of IFN-γ and IL-4 in serum was measured by ELISA. T cell subsets were detected by Immunofluorescence technique and flow cytometry. RESULTS: The content of IFN-γ and IL-4 in the serum was decreased of model group (P<0.01). The naive T cell related molecule, CD45RA was decreased(P<0.05). T cell activation-related molecule, CD25 was decreased(P<0.05). The Foxp3 in CD4(+);CD25(+); T cell was increased(P<0.01). CONCLUSION: In spleen of the ageing mice, the percentage of naive and active T cell are decreased, but The percentage of CD4(+); Tr subset is increased.