To identify the pathogenic gene and mutation site of a Chinese family with congenital cataract.Eight family members and 100 controls were employed, and targeted exome sequencing was used to identify the genetically pathogenic factor of the proband.Targeted next-generation sequencing identified a novel missense mutation c.209A>C (p.Q70P) of CRYBB1 gene in the family. Sanger sequencing results showed that this heterozygous mutation was a causative mutation, which was not found in unaffected family members and healthy controls. Bioinformatics predicts that the effect of this mutation on protein function is probably harmful.We demonstrate that c.209A>C of CRYBB1 gene is a pathogenic mutation in the family of congenital nuclear cataract in this study. This is the first report that this mutation leads to congenital nuclear cataract, which broadens the mutation spectrum of CRYBB1 gene in congenital nuclear cataract.
Acute rejection (AR) is a major complication post renal transplantation, with no widely-accepted non-invasive biomarker. This study aimed to explore the expression profiles of long non-coding RNAs (lncRNAs) in the peripheral blood (PB) of renal transplant recipients and their potential diagnostic values.The genome-wide lncRNA expression profiles were analyzed in 150 PB samples from pediatric and adult renal transplant (PRTx and ARTx) cohorts. The diagnostic performance of differentially expressed lncRNA was determined using receiver operator characteristic curve, with area under the curve (AUC) and 95% confidential interval (CI). Finally, a risk score was constructed with logistical regression model.A total of 162 lncRNAs were found differentially expressed in PRTx cohort, while 163 in ARTx cohort. Among these identified lncRNAs, 23 deregulated accordingly in both cohorts, and could distinguish AR recipients from those without AR. Finally, a risk score with two most significant lncRNAs (AF264622 and AB209021) was generated and exhibited excellent diagnostic performance in both PRTx (AUC:0.829, 95% CI:0.735-0.922) and ARTx cohorts (AUC: 0.889, 95% CI: 0.817-0.960).A molecular signature of two lncRNAs in PB could serve as a novel non-invasive biomarker for the diagnosis of AR in both pediatric and adult renal transplant recipients.
Objective To discuss the value of leukocyte-targeted myocardial contrast echocardiography (MCE) as a tool in observing the degree of acute rejection after heart transplantation. Methods Abdominal heterotopic cardiac transplantation was performed on 32 rats successfully, among which 8 isografts served as group A, and groups B, C and D involved 8 allografts respectively. The rats in groups B and C were treated with cyclosporine A (CsA) at a high dose (10mg· kg-1 · day-1 ), a low dose (3 mg · kg-1 · day-1 ) from 3rd day before transplantation respectively.The rats in groups A and D were untreated with CsA. MCE was performed during continuous intravenous SonoVue injection postoperatively on the third day after operation. We performed 2 types of MCE: perfusion imaging and leukocyte-targeted imaging. The images were obtained at 20 s and 5 min after injection of contrast agent. The value of the contrast image grayscale (GS) was measured by image analyzer (GS20s, GS5 min). GStarget was calculated as the GS5min minus the GS20s in the same rat.Postmortem histology was performed after observation. The degree of myocardial rejection was determined by HE-stained graft myocardium. Immunohistochemistry was performed to quantify the CD3-positive cells, and correlation analysis was performed between CD3-positive cell count and GS20s,GS5min, GStarget. Results Perfusion imaging showed no significant difference in myocardial GS20s of each group. Leukocyte-Targeted imaging exhibited a clear gradient in these groups (P＜0. 05). There was significant difference in GStarget of each group (P＜0. 001). Postmortem histology showed 0- Ⅰ grade rejection in group A, Ⅰ -Ⅱ grade rejection in group B, Ⅱ-Ⅲ grade rejection in group C, Ⅲ-Ⅳ grade rejection in group D. Immunohistochemistry revealed the CD3-positive cell infiltration was increased in turn from the group A to the group D. There was a significantly positive correlation between the CD3-positive cell count and GStarget ( r = 0. 86, P ＜ 0. 001 ). Conclusion Leukocyte-targeted contrast echocardiography may thus offer a noninvasive and effective ultrasound imaging technique for detecting the degree of acute cardiac transplant rejection.
Key words:
Heart transplantation; Graft rejection; Myocardium; Ultrasonography
Background: Clear cell renal cell carcinoma (ccRCC) is the most prevalent histologic subtype of kidney cancers in adults, which could be divided into two distinct subgroups according to the BRCA1 associated protein-1 (BAP1) mutation status. In the current study, we comprehensively analyzed the genome-wide microRNA (miRNA) expression profiles in ccRCC, with the aim to identify the differentially expressed miRNAs between BAP1 mutant and wild-type tumors, and generate a BAP1 mutation-specific miRNA signature for ccRCC patients with wild-type BAP1. Methods: The BAP1 mutation status and miRNA profiles in BAP1 mutant and wild-type tumors were analyzed. Subsequently, the association of the differentially expressed miRNAs with patient survival was examined, and a BAP1 mutation-specific miRNA signature was generated and examined with Kaplan-Meier survival, univariate and multivariate Cox regression analyses. Finally, the bioinformatics methods were adopted for the target prediction of selected miRNAs and functional annotation analyses. Results: A total of 350 treatment-naïve primary ccRCC patients were selected from The Cancer Genome Atlas project, among which 35 (10.0%) subjects carried mutant BAP1 and had a shorter overall survival (OS) time. Furthermore, 33 miRNAs were found to be differentially expressed between BAP1 mutant and wild-type tumors, among which 11 (miR-149, miR-29b-2, miR-182, miR-183, miR-21, miR-365-2, miR-671, miR-365-1, miR-10b, miR-139, and miR-181a-2) were significantly associated with OS in ccRCC patients with wild-type BAP1. Finally, a BAP1 mutation-specific miRNA signature consisting of 11 miRNAs was generated and validated as an independent prognostic parameter. Conclusions: In summary, our study identified a total of 33 miRNAs differentially expressed between BAP1 mutant and wild-type tumors, and generated a BAP1 mutation-specific miRNA signature including eleven miRNAs, which could serve as a novel prognostic biomarker for ccRCC patients with wild-type BAP1.
The occurrence and development of gastric adenocarcinoma (gADC) is closely related to the interaction between tumor cells and immune cells in the tumor microenvironment (TME). Our objective was to characterize the repertoire of immune cells in the TME of gADC. To analyze the transcriptomic, immune, and spatial information of TME in gADC, we constructed single-cell RNA sequencing, 10 × Genomics V(D)J analysis, multiple immunofluorescence techniques, and OSCmap analysis of 49,765 single cells in seven samples from four gADC patients. Our integrative analysis of B cells demonstrated that a large number of mucosal associated lymphoid tissue (MALT)-B cells were detected in the gADC tissues, which have mature tertiary lymphatic structures (mTLSs), and almost no MALT-B cells in peripheral blood sample. Moreover, MALT-B cells are a class of IgA+ plasma cells, which are characterized with high expression of complement pathway activation-related genes. Next, natural killer T (NKT) cells mainly exist in gADC tissues accompanied by mTLSs. This study also classified monocytes/macrophages and epithelial cells into benign and malignant types. Interestingly, CSOmap (q < .05) and multiple immunofluorescence (p < .05) results indicated more types of immune cells can be enriched in tissues with mTLSs than normal TLSs, and the density of mTLSs were higher than normal TLSs. Our findings provide novel insights for the signature of immune cells and tumor cells in the TME of gADC with TLSs and highlight the potential importance of IgA-mediated humoral immunity in gADC patients with TLSs.
Ischemic preconditioning (IPC) could protect against subsequent renal ischemia reperfusion injury (IRI). However, the mechanisms underlying IPC remain far from complete. Hence, we explored the effects of IPC on the renal and systemic hemodynamic changes, renal function and morphology, as well the involvement of endothelial and inducible nitric oxide synthase (eNOS/iNOS), and nitric oxide (NO).Male Sprague-Dawley rats were randomly divided into five groups after right-side nephrectomy: Sham group (surgery without vascular clamping); IRI group (the left renal artery was clamped for 45 min); IPC group (pretreated with 15 min of ischemia and 10 min of reperfusion); IPC + vehicle group (administrated with 0.9% saline 5 min before IPC); and IPC + N(G)-nitro-L-arginine methylester (L-NAME) group (pretreated with L-NAME 5 min prior to IPC). The renal and systemic hemodynamic parameters, renal function and morphology, as well as eNOS, iNOS, and NO expression levels in the kidneys were measured at the indicated time points after reperfusion.IPC rats exhibited significant improvements in renal function, morphology, and renal artery blood flow (RABF), without obvious influence on the systemic hemodynamics and renal vein blood flow. Increased eNOS, iNOS, and NO expression levels were detected in the kidneys of IPC rats 24 h after reperfusion. Furthermore, the beneficial effects were fully abolished by the administration of L-NAME.The results suggest that IPC contributes to early restoration of RABF, probably through eNOS/iNOS-mediated NO production, thereby alleviating the renal dysfunction and histological damage caused by IRI.
Objective
To investigate the relationship between blood vessel invasion (BVI) and clinicopathologic features and prognosis in patients with gastric cancer, and analyze related factors affecting the prognosis of patients.
Methods
The clinicopathological data of 206 patients with gastric cancer who were admitted to the Nanjing Hospital Affiliated to Nanjing Medical University from January 2007 to December 2010 were retrospectively analyzed. The BVI of surgical tumor specimens in all patients was detected by immunohistochemical staininng. All the patients were followed up via outpatient examination and telephone interview up to March 2014. The count data were analyzed using the chi-square test. The survival curve was drawn by Kaplan-Meier method. The survival analysis and univariate analysis were done using the Log-rank test, and multivariate analysis was done using the COX regression model.
Results
The BVI rate of 206 patients was 27.67% (57/ 206). The BVI rate of tumor tissues, tumor differentiation, perineural invasion, T stage, N stage and TNM stage in all patients with gastric cancer were compared, showing significant differences (χ2 =14.396, 9.569, 15.579, 43.453, 30.732, P <0.05). After operation, 188 patients were followed up for 6.0-60.0 months (median, 34.0 months) , with the follow-up rate of 91.26% (188/ 206). Among 188 patients with follow-up, the median survival time and 5-year cumulative survival rate in patients with BVI and with negative BVI were 32.4 months and 19.6% , 40.7 months and 42.0% , respectively, with a significant difference in the survival of patients (χ2 =9.364, P <0.05). The results of univariate analysis showed that the diameter of tumor, tumor differentiation, perineural invasion, BVI, T stage, N stage and TNM stage were factors affecting the prognosis of patients with gastric cancer (χ2=9.241, 17.486, 11.243, 9.364, 27.666, 216.745, 49.887, P <0.05). The results of multivariate analysis showed that the diameter of tumor more than 5 cm, BVI, stage T4 , stage N3 and stage Ⅲ were independent risk factors affecting the prognosis of patients with gastric cancer (HR =0.502, 0.456, 0.052, 0.001, 0.735; 95% confidence interval: 0.334-0.754, 0.289-0.720, 0.004-0.664, 0.000-0.006, 0.159-3.398, P <0.05).
Conclusions
BVI in patients with gastric cancer is associated with the progression of tumors. The diameter of tumor more than 5cm, BVI, stage T4 , stage N3 and stage Ⅲ are independent risk factors affecting the prognosis of patients with gastric cancer, and BVI may be a predictor of poor prognosis of patients with gastric cancer.
Key words:
Gastric neoplasms; Blood vessel invasion; Prognosis
Abstract The roles of tumor infiltrating B lymphocytes (TIBs) and tertiary lymphoid structures (TLSs) in solid tumor genesis and anti-tumor therapy have been recognized by researchers, but the specific formation and effect of TLSs have not been fully understood. In this study, single-cell RNA sequencing, multiple immunofluorescence assays, and cellular spatial organization mapper analysis were used to explore the structures of TLSs in gastric adenocarcinoma (GADC). Furthermore, the relationships between TLSs and clinicopathological characteristics and prognosis of GADC patients were analyzed. The results identified the phenotype of immunoglobulin A (IgA) + -TLSs which contained higher level of IgA + plasma cells in GADC, and several types of immune cells in IgA + -TLSs had higher levels of cellular interactions and migration ability. Furthermore, IgA + -TLSs correlated with age, differentiation, distant metastasis, TNM stage, expression of programmed death-ligand 1, J-chain, SSR4, and good overall survival. In conclusion, IgA + plasma cells higher expressed in IgA + -TLSs, and the formation of IgA + -TLSs promotes a better prognosis for GADC patients.
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