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Objective To investigate the clinicopathological features of recurrent and de novo focal segmental glomerulosclerosis (FSGS) after kidney transplantation. Methods Thirty-four recipients pathologically diagnosed with FSGS by renal allograft biopsy were enrolled in this clinical trial. According to the detection of primary diseases of renal allografts and circulating permeability factors, 34 recipients were divided into the recurrent FSGS group (n=12) and de novo FSGS group (n=22). The differences of clinical indexes and the degree of pathological injury of renal allografts were compared between two groups. Results There was no significant difference in the mesangial hyperplasia score, glomerulosclerosis rate, renal tubular atrophy score, interstitial fibrosis score and podocyte proliferation rate between two groups (all P > 0.05). In the recurrent FSGS group, segmental glomerulosclerosis rate of the recipients was 0.10 (0.08, 0.27), lower than 0.19 (0.13, 0.33) in the de novo FSGS group (P < 0.05). No significant difference was found in the incidence of antibody-mediated rejection, drug-induced renal tubular injury and BK virus infection between two groups (all P > 0.05). The incidence of T cell-mediated rejection in the recurrent FSGS group was 17%, lower than 55% in the de novo FSGS group (P < 0.05). Immunohistochemical staining showed that the infiltrating inflammatory cells in the renal allografts were mainly T lymphocytes. The positive rates of C4d deposition in peripheral capillaries between the recurrent and de novo FSGS groups were 33% (4/12) and 32% (7/22), with no significant difference (P > 0.05). Immunofluorescence results revealed IgM deposition in the segmental glomerulosclerosis area of renal allografts in most cases. Electron microscopy showed extensive fusion or segmental distribution of podocytes in the glomerulus of renal allografts. Conclusions The degree of renal functional injury and the incidence of T cell-mediated rejection in the recurrent FSGS group are lower than those in the de novo FSGS group. Comprehensive analysis of preoperative and postoperative clinical manifestations, laboratory testing and pathological examination of kidney transplant recipients contribute to early diagnosis and treatment of recurrent and de novo FSGS.
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Objective:To determine the accuracy and clinical application of donor HLA quartile genotyping based upon transplanted kidney biopsy.Methods:The clinical and follow-up data are retrospectively reviewed for 38 recipients of kidney transplantation(KT)at First Affiliated Hospital of Xi'an Jiaotong University from 2019 to 2022.They are suspected of rejection.HLA quartile genotyping of donor kidney is performed through puncture and DNA analysis by LABType SSO method.Known HLA genotypes of recipients are compared for predicting HLA genotypes of donors.Donor-specific antibody(DSA)is detected by Labscreen Single kit.And SPSS18.0 statistical software is employed for processing baseline data, donor/recipient HLA typing data, recipient DSA antibody data and transplant nephropathy parameters.Results:Among them, 12(31.58%)belonged to HLA-A, B, C, DR and DQ.Four loci are detected in 14 cases(36.84%). Three sites are detected in 10 cases(26.32%). Two sites are detected in 2 cases(5.26%)and a negative correlation exists between detected sites and transplantation time( rs=-0.707, P=0.001). The detection rate of HLA loci is 78.94%(30 cases). B: 65.78%(25 cases); C: 84.21%(32 cases); DR: 57.89%(22 cases); DQ: 100% (38 cases); HLA sites detected in puncture tissue are 89.47% consistent with the results of donor whole blood test, among which HLA-C and HLA-DQ sites are 100% consistent and HLA-A and HLA-B sites 87.50% and 90% consistent and HLA-DR sites 66.7% consistent( P<0.01). Spearman's rank correlation analysis indicated that pathological diagnosis of interstitial inflammation( rs=-0.432, P=0.017), renal tubule atrophy( rs=-0.587, P=0.001)and interstitial fibrosis( rs=-0.560, P=0.001)are correlated negatively with HLA detected sites in transplanted kidney puncture tissue.DSA is detected in 42.1% of recipients and 68.75% of recipients belonged to HLA-DQ. Conclusions:HLA typing results of puncture tissue are consistent with those of whole blood test.Time after transplantation, infiltration of transplanted nephritis cells and degree of fibrosis may influence the detection of HLA loci.Donor HLA quartile genotyping using transplanted kidney biopsy has some diagnostic values for detecting the presence of DSA.
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Objective:To investigate the gene transcription level changes in the amygdala of social isolation rearing models of schizophrenia to determine the pathogenic genes and their related pathways of schizophrenia.Methods:A total of 29 3-week-old SPF C57BL/6J male mice were randomly divided into control group ( n=16) and model group ( n=13); 4 mice were raised in each transparent mouse cage in the control group, and 1 mouse was raised in each transparent mouse cage in the model group; mice in each cage could see their surrounding mice but could not touch each other. Mice in both groups were fed for 4 weeks and then subjected to open field experiment, pre-pulse inhibition experiment and new object recognition experiment within one week. After the experiment, mice were sacrificed by spinal dislocation, and the amygdala was taken for transcriptome sequencing. The topGO software was used for gene ontology (GO) functional enrichment analysis of differentially expressed genes (DEGs), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed using KEGG database. Results:(1) Animal experiment: compared with the control group, the model group had significantly increased movement distance in the open field experiment ([1 239.20±106.35] m vs. [1 845.53±143.65] m, t=3.464, P=0.002), significantly decreased activity time in the central region 5 min before experiment ([13.15±1.41] s vs. [8.47±1.19]) s, t=2.464, P=0.020). Compared with the control group, the model group had significantly lower percentage of deficient prepulse inhibition (PPI) of 78 dB ([22.28±1.53] % vs. [14.59±2.75] %, t=2.629, P=0.013), and deficient PPI of 88 dB ([32.83±3.39] % vs. [18.44±3.07] %, t=3.081, P=0.005). Compared with the control group, the model group had significantly decreased ratio of time exploring new objects/time exploring former objects ([80.5±2.2]% vs. [71.0±3.6]%, t=2.356, P=0.026). (2) Bioinformatics analysis: a total of 96 DEGs were found, of which 42 were with up-regulated expressions ( Th, Crlf1, etc.), and 54 were with down-regulated expressions ( Prkcd, etc.). Th and Crlf1 were positively correlated ( r=0.940, P=0.018). GO enrichment results suggested that DEGs were enriched in projection function of plasma membrane boundary cells, neuronal differentiation, and cell apoptosis. KEGG enrichment results suggested that DEGs were enriched in WNT signaling pathway, apoptosis pathway and tyrosine metabolism pathway. Protein network interaction analysis suggested that Wnt6, Tcf712, Pitx2, Tcf7 and Cd4 were key proteins. Conclusion:DEGs such as Th, Prkcd, Lrrc74b, Fadd, Wnt6, Ror2, Notum, and Tcf7l2, and their related signaling pathways may be related to schizophrenia in the amygdala of social isolation rearing mice.
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Objective: FF456 is a new gene cloned in our lab, which belongs to c-type G protein-coupled receptors and has high homology to the olfactory receptor family. Northern blotting and RT-PCR showed that the expression of FF456 was exclusively restricted in liver and was significantly down-regulated in hepatoma. In an effort to study the function of FF456 gene, high titer antibody is indispensable. Methods:Full-length cDNA of FF456 was reconstructed into pcDNA3.1(-) vector, which was used for immunizing 6- to 8- weeks old female BALB/c mice. The effects of different injection manners, solvents and dosages on the antibody production have been compared. For detecting the titer and specificity of the antisera produced by DNA immunization, a peptide containing FF456 specific sequence was expressed by E.coli expression system. Results:Finally specific antisera with titers as high as 1 ∶ 50 000 was obtained. Conclusion:Immunofluorescence assays showed FF456 was expressed in hepatocytes with high tissue specificity. The FF456 expression in hepatoma cells was decreased dramatically.