1.Construction, breeding, and gene identification of TREM2 knockout mice
Rong Huang ; Xinxin Zhao ; Hui Xue ; Mengjuan Zhu ; Jiajie Tu ; Xinming Wang
Acta Universitatis Medicinalis Anhui 2025;60(6):977-983
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.
2.The value of biparametric MRI in the detection of prostate cancer
Yueyue ZHANG ; Wenlu ZHAO ; Chaogang WEI ; Tong CHEN ; Mengjuan LI ; Shuo YANG ; Shuangxiu TAN ; Beibei HU ; Qi MA ; Yongsheng ZHANG ; Boxin XUE ; Junkang SHEN
Chinese Journal of Radiology 2019;53(2):109-114
Objective To explore the difference in efficacy between multiparametric MRI (Mp-MRI) based on prostate imaging reporting and data system version 2 (PI-RADS v2) and abbreviated biparametric MRI (Bp-MRI) in detecting prostate cancer (PCa) and clinically significant prostate cancer (csPCa), and to evaluate the consistency of image interpretation between different readers. Methods The imaging, pathological and clinical data of patients with prostatic Mp-MRI in our hospital from February 2015 to June 2018 were retrospectively analyzed. At the beginning, 250 patients were randomly selected. Two radiologists visually evaluated the images of those patients using two 5-point scoring schemes based on Mp-MRI and Bp-MRI. The remaining cases were independently proceeded by one of the radiologists using two schemes respectively. Weighted Kappa test was used to assess the consistency of the results interpreted by the two radiologists. The receiver operating characteristic (ROC) curve was used to evaluate the efficiency of the two scoring schemes in detecting PCa and csPCa, and with Z test to investigate whether there was any difference in detection efficiency between the two schemes. Results Nine hundred and seventy eight patients were eventually enrolled in the study. The results of the consistency assessment showed that there was good agreement between the two radiologists, whether using Mp-MRI or Bp-MRI, with the weighted Kappa coefficient of 0.800 and 0.812, respectively. The ROC curve analysis showed that the area under the curve (AUC) of PCa detected by Mp-MRI and Bp-MRI was 0.873 and 0.879, respectively, and the AUC of csPCa detected was 0.922 and 0.932, respectively. In addition, there was no statistically significant difference between the AUC of PCa and csPCa detected by the two schemes (P>0.05). Conclusion The Bp-MRI scoring scheme has good stability in the evaluation of benign and malignant prostate, and its detection efficiency of PCa or csPCa is not lower than that of standard Mp-MRI based on PI-RADS v2.
3.Influence of interleukin-2 receptor antagonists on the morbidity and prognosis of new-onset diabetes after liver transplantation
Jing LIANG ; Mengjuan XUE ; Xianying CHEN ; Xiaowu HUANG ; Qiman SUN ; Ting WANG ; Jian GAO ; Jian ZHOU ; Jia FAN ; Mingxiang YU
Chinese Journal of Endocrinology and Metabolism 2018;34(2):121-128
Objective To explore the influence of interleukin-2 receptor antagonists(IL-2Ra) on the morbidity and prognosis of new onset diabetes after transplantation(NODAT)in liver transplant recipients. Methods Pre-and post-operative clinical data of 879 nondiabetic patients who underwent a liver transplantation between April 2001 and December 2016 were retrospectively studied. All the enrolled patients were divided into IL-2Ra and non-IL-2Ra groups according to the use of IL-2Ra. Transient-NODAT(T-NODAT)and Persistent-NODAT(P-NODAT)were defined according to whether NODAT would be existed continuously. The impacts of IL-2Ra on the cumulative incidence as well as the risk of NODAT and T-NODAT were analyzed through comparison between patients who used IL-2Ra or not. And influence of IL-2Ra on the long-term survival of NODAT patients was further analyzed. Results Among 879 patients,177(32.24%)from the IL-2Ra group(n=549)developed NODAT and 29.38%(n=52)of the NODAT reversed,while 131(39.70%)from the non-IL-2Ra group(n=330)developed NODAT and 26.72%(n=35)of the NODAT reversed. After adjusting for 18 possible confounding factors,the IL-2Ra group had significantly decreased cumulative incidence of NODAT over the non-IL-2Ra group(adjusted P=0.028). COX regression analyses showed that IL-2Ra was a protective factor against NODAT development(HR 0.774;95% CI 0.616-0.973; P=0.028), while the use of IL-2Ra and the reverse of NODAT did not significantly related. In addition, long-term survival of the NODAT patients were far better in the IL-2Ra group(adjusted P=0.001). Conclusion IL-2Ra significantly reduces the risk of NODAT in liver transplant recipients and is beneficial to the long-term survival of NODAT patients.
4.The analysis of bone metabolism in hospitalized patients with Graves disease and the changes after 131I therapy
Wenjie MA ; Chaoyang LV ; Yao ZHANG ; Guiping XU ; Shunmei HE ; Mengjuan XUE ; Jian GAO ; Mingxiang YU
Fudan University Journal of Medical Sciences 2017;44(2):186-191
Objective To analyze the bone metabolism in hospitalized patients with Graves disease and the changes after 131I therapy.Methods The differences of bone metabolism were analyzed between 315 patients with Graves disease and 300 normal controls in a case-control study.The changes in bone turnover markers and BMD levels before and one year after 131I therapy were observed in 60 patients.Results Compared to normal control,bone turnover markers were markly higher and BMD levels were lower in patients with Graves disease.The level of thyroid hormones were positively related to bone turnover markers,while negatively related to total hip BMD (Z-score).But there was no linear relationship with lumbarand femoral neck BMD (Z-score).After one year of 131I therapy,bone turnover markers were markly lower than that before treatment,while BMD levels were partly higher than that before treatment.Conclusions In Graves disease patients,bone turnover markers were generally increased,while BMD levels decreased compared with normal people.After 131I therapy,along with the improvement of thyrotoxicosis,the high bone turnover rate can be suppressed,and BMD can partly recover.
5.Effects of different anti-rejection drugs and projects on dyslipidemia after organ transplantation
Mengjuan XUE ; Chaoyang LYU ; Yao ZHANG ; Shunmei HE ; Mingxiang YU
Chinese Journal of Endocrinology and Metabolism 2015;(9):816-818
[Summary] Dyslipidemia after organ transplantation is one of the important risk factors of postoperative cardiovascular disease and graft dysfunction. There are many factors that result in postoperative dyslipidemia. However, the factors influencing serum lipid levels are changing with the development of organ transplantation. In this article the effects of different anti-rejection drugs such as cyclosporine, azathioprine, mycophenolate mofetil, tacrolimus, rapamycin ( sirolimus ) , corticosteroids, and monoclonal antibody on dyslipidemia after organ transplantation were summarized in different eras.


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