1.Expression of miR-151a-3p in peripheral blood mononuclear cells of children with lupus nephritis and its correlation with disease activity and vascular damage
Hui YANG ; Qiuyu LIN ; Yongzhao CHEN ; Kaihui XING
Chinese Journal of Applied Clinical Pediatrics 2021;36(5):359-363
Objective:To investigate the expression of miR-151a-3p in peripheral blood mononuclear cells (PBMCs) of children with lupus nephritis (LN) and its correlation with activity and vascular damage.Methods:A total of 87 children with LN admitted in Hainan Women and Children′s Medical Center from January 2016 to March 2019 were enrolled, including 16 cases of type Ⅱ, 14 cases of type Ⅲ, 34 cases of type Ⅳ, 17 cases of type Ⅴ and 6 cases of type Ⅵ.Besides, additional 40 children with normal physical examination were selected as the healthy control group.The 87 children with LN were divided into the LN stable group (31 cases) and LN active group (56 cases) by using systemic lupus erythematosus disease activity index (SLEDAI) scores.According to different proportions of the medium membrane, the patients were classified into the mild group (25 cases), moderate group (34 cases) and severe group (28 cases). Real time quantitative PCR (qPCR) was used to detect the expression level of miR-151a-3p in PBMCs of each group.The correlation of the expression level of miR-151a-3p in PBMCs of LN children with the SLEDAI score and renal interstitial damage score was studied by Pearson correlation analysis. Results:The expression levels of miR-151a-3p in the LN group, LN active group and LN stable group (0.47 ± 0.06, 0.30 ± 0.04, and 0.75±0.12, respectively) were significantly lower than that in the healthy control group (1.62±0.35) (all P<0.01), and the expression level of miR-151a-3p in the LN active group (0.30±0.04) was significantly lower than that in the LN stable group (0.75±0.12) ( P< 0.01). The SLEDAI score [(14.35±4.60) points vs.(8.25±2.24) points] and renal interstitial damage score [(52.70±6.30) points vs.(34.80±4.10) points] in the LN active group were significantly higher than those in the LN stable group (all P<0.01). The expression levels of miR-151a-3p (0.18±0.02, 0.41±0.05 vs.0.83±0.16) in type Ⅴ-Ⅵ and type Ⅳ groups were significantly lower than that in the type Ⅱ-Ⅲ group (all P<0.01). The SLEDAI scores [(16.50±5.28) points, (12.36±3.74) points vs.(6.40±1.70) points] and renal interstitial damage scores [(62.70±7.40) points, (47.20±5.80) points vs.(25.80±3.50) points] in type Ⅴ-Ⅵ and type Ⅳ groups were significantly higher than those in the type Ⅱ-Ⅲ group (all P< 0.01). The expression levels of miR-151a-3p (0.20±0.02, 0.39±0.04, 0.86±0.18 vs.1.62±0.35) in severe, moderate and mild groups were significantly lower than that in the healthy control group (all P<0.01), and the expression level of miR-151a-3p decreased with the aggravation of vascular damage.The SLEDAI scores [(15.20±5.10) points, (12.85±3.90) points vs.(6.70±1.82) points] and renal interstitial damage scores [(57.30±6.80) points, (51.60±6.30) points vs.(27.20±3.60) points] in the severe and moderate groups were significantly higher than those in the mild group (all P<0.01). The expression level of miR-151a-3p in LN children was negatively correlated with the SLEDAI score and renal interstitial damage score ( r=-0.682, -0.627, all P<0.001). Conclusions:The expression level of miR-151a-3p in PBMCs of LN children is significantly decreased.The declined miR-151a-3p level is closely related to disease activity and vascular damage, and may be involved in the occurrence and development of LN.
2.Preparation and cytocompatibility of beta-calcium phosphate scaffolds with tanshinol coating
Tao WU ; Yingchao LIU ; Kaihui NAN ; Tao SHU ; Jinhui WU ; Zhiqin LU ; Jianzhou XING ; Jun LV
Chinese Journal of Tissue Engineering Research 2017;21(14):2247-2253
BACKGROUND: Revascularization is a challenge for the tissue-engineered bone carrying cells after implanted into human body. Previous studies have found that tanshinol can improve the functions of endothelial progenitor cells and exert vascular protective effects.OBJECTIVE: To prepare the β-calcium phosphate (β-TCP) scaffold with tanshinol coating, and to observe its cytocompatibility.METHODS: The β-TCP scaffolds coated with 10-7, 10-6 and 10-5 mol of tanshinol were constructed by negative pressure absorption method. The distribution of tanshinol coating on the scaffold was observed using scanning electron microscopy,and the inner ingredients were analyzed by infrared spectrum. Human endothelial progenitor cells (hEPCs) were cultured in the extracts of β-TCP and β-TCP scaffolds with 10-7, 10-6, 10-5 mol of tanshinol coatings, respectively. The cell proliferation was detected at 2, 4, 6, 8 and 10 days of culture; the levels of nitric oxide and vascular endothelial growth factor in the supernatants were detected at 1, 7 and 14 days of culture; the lumen formation on the matrigel was observed after 14-day culture. hEPCs were respectively seeded onto the β-TCP and β-TCP scaffolds with different dosages of tanshinol coating,and then the cell growth was observed under scanning electron microscope at 7 days.RESULTS AND CONCLUSION: The tanshinol coating evenly distributed on the inner surface of the pores, and its crystalline structure became dense with dosage increasing. Infrared spectrum analysis revealed no changes in the characteristic absorption peak of tanshinol and TCP in the scaffold. The β-TCP scaffolds with tanshinol coating could promote the proliferation of hEPCs, especially the scaffolds with 10-6 and 10-5 mol tanshinol coating. Compared with the β-TCP scaffold, the scaffolds with 10-6 and 10-5 mol tanshinol coating significantly upregulated the nitric oxide level at 14 days of culture, and significantly increased the level of vascular endothelial growth factor at 7 and 14 days of culture (P <0.05). Although it could be found in all β-TCP scaffolds with tanshinol coating, the lumen formation was the maturest in the scaffold with 10-5 mol tanshinol coating. These results suggest the β-TCP scaffolds with tanshinol coating can promote the proliferation and endothelial differentiation of hEPCs, and hold a good cytocompatibility.
3.Clinical phenotype and genetic analysis of a Chinese patient featuring X-linked Claes-Jensen type syndromic mental retardation.
Min GAO ; Mengjuan XING ; Kaihui ZHANG ; Yuqiang LYU ; Jian MA ; Zhongtao GAI ; Yi LIU
Chinese Journal of Medical Genetics 2020;37(7):736-738
OBJECTIVE:
To explore the genetic basis of a Chinese patient featuring global developmental delay.
METHODS:
Peripheral venous blood samples from the proband and his parents and sister were taken for the extraction of DNA. Target capture and next generation sequencing was carried out to detect genetic variants associated with the disease. Suspected variant was validated by Sanger sequencing.
RESULTS:
Genetic testing discovered that the proband has carried hemizygous c.150G>T and c.150+1G>T variants of the KDM5C gene which are inherited from his mother. His younger sister also carried the variants. The c.150+1G>A variant was unreported previously, which has altered a splice site and was predicted to be pathogenic by bioinformatics analysis.
CONCLUSION
The hemizygous c.150+1G>T variant of the KDM5C gene, known to underlie X-linked Claes-Jensen type syndromic mental retardation, probably accounts for the disorder in the patient. Identification of this variant has enriched the variant spectrum of the KDM5C gene.