Objective To develop a candidate reference method for the determination of serum creatinine and to evaluate the ac-curacy of conventional detection systems though method comparison to achieve traceability .Methods The candidate reference method was established according to the sarcosine oxidase and the accuracy and reliability of the method was verified through par-ticipation in international reference laboratories EQA activities (IFCC-RELA) .20 fresh single human serum samples with different concentration and calibrator were simultaneously measured by using conventional detection system and candidate reference method . Results The calibration curve for serum creatinine was linear in the concentration range from 50-2 000 μmol/L with a correlation coefficient of 0 .999 9 under the optimum experimental conditions (the linear equation was Y=0 .000 884 2X-0 .000 325 3) and the imprecision was less than 1 .0% .The proposed method has been applied to the determination of RELA samples with satisfactory re-sults .The measured results with conventional detection systems were consistent with candidate reference method ,and the slope of the regression equation was 1 .005 6 .Conclusion The candidate reference method of serum creatinine is successfully established and which can be used for traceability and standardization .It may provide an effective way for conventional detection system traceable to the reference method or reference material .

Objective To establish a candidate reference method for the determination of serum lithium based on ion chromatog-raphy and evaluate its analytical performance.Methods A simple sample treatment procedure,which can be remove the proteins and/or organics in human serum,has been developed for the determination of serum lithium.Method precision was evaluated with different concentration of fresh human serum and EQA sample RELA-A/B.Method accuracy was investigated with the recovery ex-periments in fresh human serum and RELA-A/B sample.Results The linear equation was Y =0.817 1X -0.001 3 with a correla-tion coefficient of 0.999 95 under the optimum experimental conditions,the detection limit (3S/N)for lithium was 6 μg/L and the imprecision was less than 1.0%.The results of the recovery experiments indicated that the recoveries were reasonable for the deter-mination of serum lithium,in a range of 99%-101%.Conclusion The candidate reference method of lithium was successfully es-tablished and which can be used for traceability and standardization.It may provide an effective way for routine testing of lithium traceable to the reference method/reference material.

Objective To investigate the application value of urine microalbumin (mALB) ,retinol binding protein(RBP) and cys-tatin C(CysC) and their combined detection in early diagnosis of type 2 diabetic nephropathy(DN) .Methods Ninety-two inpatients with DN (DN group) and 90 people undergoing the physical examination(control group) in our hospital from June 2014 to Decem-ber 2015 were collected .Urine mALB ,RBP and CysC were detected in all subjects and detection results were analyzed statistically . Results The levels of urine mALB ,RBP and CysC in the DN group were significantly higher than those in the control group ,the differences all had statistical significance (P< 0 .05) .Among 3 indicators ,the positive rate of urine mALB for detecting DN was highest (94 .57% ) ,while which of 3-index combined detection was 97 .83% ,and significantly higher than that of single detection , the difference was statistically significant(P<0 .05) .The sensitivity ,specificity ,positive predictive value ,negative predictive value and Youden index of 3-index combined detection were all higher than those of single index .The ROC curve showed that AUC of u-rine mALB for diagnosing DN was 0 .732 ,the diagnostic cut-off value was 43 .58 mg/L ,AUC of urine RBP was 0 .685 ,the diagnos-tic cut-off value was 1 .47 mg/mL ,AUC of urine CysC was 0 .701 ,the diagnostic cut-off value was 1 .42 mg/L ,while AUC of com-bined detection was 0 .928 .Conclusion Urine mALB ,RBP and CysC are better indexes reflecting renal injury .Their combined de-tection will increase the positive rate ,sensitivity and specificity for diagnosing DN .So monitoring the levels of urine mALB ,RBP and CysC has an important significance to diagnosing the occurrence and development of DN early renal injury and prevention ,treat-ment and delaying progress of DN .

The spatial structure of hemolymph node in the rat is studied by light, transmission and scanning electron microscopy after the fixation of arterial perfusion. The structure of hemolymph node is similar to that of the normal lymph node, and main characteristic is that a number of the red cells are seen in it. The erythrocytes were carried to the afferent lymphatic vessel and reach the medullary sinus, many erythrocytes travel through the rsubcapsula and cortical sinuses, and reach the lymphatic tissue of the local cortex with selectivity, and going through the paracortical zone and the sinus wall to the medulary sinus. Most of the red blood cells are phagocytosed by macrophages in the sinuses. The subcapsular and cortical sinuses of hemolymph nodes connect with medullary sinuses, and form a reticular lymphatic passage. Reticular cells in the sinuses constitute a spatial each other. There are macrophages, lymphocytes, plasmocytes and numerous red blood cells in the nets. Macrophages are anchored on the reticular cells by their pseudopodla, traping and phagocytosing the red cells and foreign matters. Sometimes a macrophage is found closely associated with lymphocytes.

Aim To explore the possibility of the multiple epitope DNA vaccines of hepatitis C virus (HCV). Methods A synthetic multiple epitope antigen gene PCX of HCV was cloned into vector pREP9(RSV promoter) and pcDNA3 (CMV promoter) to construct eukaryotic expression vectors pREP9/PCX and pcDNA3/PCX, then they were used to immunize mice and rabbits, the titer of specific humoral and cellular responses were detected and their safety were observed. Results In mice, specific anti-GZ-PCX antibody(IgG) was lower than 1∶ 1 000 and did not persist well. In rabbits, the highest titer of anti-GZ-PCX IgG reached at 1∶ 3 200 and remained for about one month. Delayed type hypersensitivity reactions (DTH)and proliferation response of peripheral lymphocytes were induced by GZ-PCX antigen. Body weights of immunized mice were normal and no obvious toxic reaction was observed. Conclusion The multiple epitope antigen gene of HCV could induce specific immune responses without obvious toxicity and it might be able to serve as an effective HCV vaccine candidate.

Infectious bronchitis virus (IBV) poses a severe threat to the poultry industry and causes heavy economic losses worldwide. Vaccination is the most effective method of preventing infection and controlling the spread of IBV, but currently available inactivated and attenuated virus vaccines have some disadvantages. We developed a chimeric virus-like particle (VLP)-based candidate vaccine for IBV protection. The chimeric VLP was composed of matrix 1 protein from avian influenza H5N1 virus and a fusion protein neuraminidase (NA)/spike 1 (S1) that was generated by fusing IBV S1 protein to the cytoplasmic and transmembrane domains of NA protein of avian influenza H5N1 virus. The chimeric VLPs elicited significantly higher S1-specific antibody responses in intramuscularly immunized mice and chickens than inactivated IBV viruses. Furthermore, the chimeric VLPs induced significantly higher neutralization antibody levels than inactivated H120 virus in SPF chickens. Finally, the chimeric VLPs induced significantly higher IL-4 production in mice. These results demonstrate that chimeric VLPs have the potential for use in vaccines against IBV infection.
Animals ; Antibodies, Viral/blood ; *Chickens ; Chimera/genetics/immunology ; Coronavirus Infections/prevention & control/*veterinary/virology ; Female ; *Immunity, Innate ; Infectious bronchitis virus/genetics/*immunology ; Influenza A Virus, H5N1 Subtype/genetics/immunology ; Injections, Intramuscular/veterinary ; Mice ; Mice, Inbred BALB C ; Neuraminidase/genetics ; Poultry Diseases/*prevention & control/virology ; Recombinant Fusion Proteins/genetics/immunology ; Spike Glycoprotein, Coronavirus/genetics/*immunology ; Vaccines, Synthetic/administration & dosage/genetics/immunology ; Vaccines, Virus-Like Particle/administration & dosage/genetics/*immunology ; Viral Proteins/genetics

METTL3 and METTL14 are two components that form the core heterodimer of the main RNA m6A methyltransferase complex (MTC) that installs m6A. Surprisingly, depletion of METTL3 or METTL14 displayed distinct effects on stemness maintenance of mouse embryonic stem cell (mESC). While comparable global hypo-methylation in RNA m6A was observed in Mettl3 or Mettl14 knockout mESCs, respectively. Mettl14 knockout led to a globally decreased nascent RNA synthesis, whereas Mettl3 depletion resulted in transcription upregulation, suggesting that METTL14 might possess an m6A-independent role in gene regulation. We found that METTL14 colocalizes with the repressive H3K27me3 modification. Mechanistically, METTL14, but not METTL3, binds H3K27me3 and recruits KDM6B to induce H3K27me3 demethylation independent of METTL3. Depletion of METTL14 thus led to a global increase in H3K27me3 level along with a global gene suppression. The effects of METTL14 on regulation of H3K27me3 is essential for the transition from self-renewal to differentiation of mESCs. This work reveals a regulatory mechanism on heterochromatin by METTL14 in a manner distinct from METTL3 and independently of m6A, and critically impacts transcriptional regulation, stemness maintenance, and differentiation of mESCs.
Animals ; Mice ; Methylation ; Chromatin ; Histones/metabolism* ; RNA, Messenger/genetics* ; Methyltransferases/metabolism* ; RNA/metabolism*