Objective : To construct triggering receptor expressed on myeloid cells 2(TREM2) gene knockout(TREM2-/-) mice using CRISPR/Cas9 technology, to breed TREM2-/- mice and to analyze the genotype of TREM2-/- mice. Methods : CRISPR/Cas9 technology was used to selectively knock out exon 2-3 regions of TREM2 gene to construct a TREM2-/- mouse model, and the genetic background of all mice was C57BL/6J. Polymerase chain reaction(PCR) was used to identify the genotype of mice. Quantitative real-time PCR(qPCR) and Western blot were used to detect the expression level of TREM2 in major tissues of mice, and the authenticity and scientific nature of PCR identification results were verified from mRNA level and protein level. According to the sgRNA sequence, the possible off-target sites were predicted on the CCTop website, and the tail DNA of mice was extracted and amplified by PCR and then Sanger sequencing was performed to detect whether there was off-target effect in TREM2-/- mice. Results : TREM2-/- mice were successfully constructed by CRISPR/Cas9 technology, and the mice were genotyped. The results of agarose gel electrophoresis showed that the mouse genotype with only 415 bp band amplified was wild type(WT), the mouse genotype of the 449 bp band amplified only was TREM2-/-, and the mouse genotypes amplified with 415 bp and 449 bp double bands were heterozygous. qPCR results showed that compared with WT mice, the mRNA expression of TREM2 in heart and brain tissues of TREM2-/- mice was down-regulated(P-/- mice was reduced(P-/- mice. Conclusion TREM2-/- mice are successfully constructed and bred, a reliable genotype identification method is established, the genetic stability of the mouse model is verified, which will provide an important genetic animal model for the study of TREM2 gene function.

Objective:to investigate clinical significance of serum and urine NGAL (Neutrophil gelatinase-associated lipocalin) in the early kidney damage with primary hypertension.Methods:According to UAER ( urinary microalbumin excreting rate ) ,we divided 90 patients with primary hypertension into three groups (<30 mg/24 h,30-300 mg/24 h and >300 mg/24h),and selected 30 healthy people as the control group.Serum and urine NGAL and cystatin C ,serum creatinine,urea nitrogen,high sensitive C reactive protein , transferrin,and basis of blood pressure were detected and followed up one year.Results:Compared with healthy group ,GFR( glomerular filtration rate ) and serum NGAL were decreased significantly in medium and severe proteinuria groups , while urine NGAL was increased.Conclusion:Serum and urine NGAL have been a clear trend changes in kidney damage ,which could be used as a reliable indicator in monitoring renal function of patients with primary hypertension.

Objective Severe acute respiratory syndrome is recently emerged infectious disease caused by a novel coronavirus, but its immmunopathological mechanism has not yet been fully elucidated. Method We established the SARS animel model and investigated changes in plasma inflammatoy cytokines monkeys and rats. 8 monkeys with PCR and antibody positive were detected. Serum levels of inflammatory cytokines interleukin(IL)-6,IL-10,Th-1 cytokine,interferon(INF)-? and tumor necrosis factor(TNF)-? were measured by enzyme-linked immunosorbent assays(ELISA).Result The concentration of IL-10 and TNF-?were not significantly different in model and control group.IL-6 showed marked elevation of at least 10 days, and there was a positive relationship between the level of IL-6 and pulmonary pathological changes. The INF-? level decreased. Conclusion The result of sera level of SARS animal model avoid the disturbance of anti-viral drug and corticosteroid, it suggest that there are different immunoregulatory events during SARS and may contribute to our understanding of the pathogenesis.