彩色多普勒超声引导下经胫/腓静脉-腘静脉顺行置管溶栓治疗急性混合型下肢深静脉血栓形成的疗效观察

目的 探讨彩色多普勒超声引导下经小腿胫/腓静脉-腘静脉顺行置管溶栓治疗急性混合型下肢深静脉血栓形成（DVT）的疗效。方法 横断面研究。纳入2019年8月—2022年8月江苏大学附属武进医院介入血管科经下肢静脉造影确诊为急性下肢DVT累及腘静脉的患者27例。其中男8例、女19例,年龄23~75（55.3±14.3）岁。彩色超声多普勒引导下穿刺小腿胫前、胫后或腓静脉,顺行置入溶栓导管,溶栓段覆盖血栓全程,在充分抗凝基础上间断泵入尿激酶,每24~48 h经导管行造影复查,并调整溶栓导管位置。观察指标：彩色多普勒超声引导下经小腿胫/腓静脉-腘静脉顺行置管穿刺成功率及并发症发生情况;溶栓前后健侧与患侧肢体周径差、患肢静脉总通畅度评分和溶栓后通畅度,以及腘静脉通畅度评分和溶栓后通畅度。结果 27例患者经小腿胫/腓静脉-腘静脉顺行置管成功,其中彩色超声多普勒导引下穿刺成功22例（穿刺成功率为81.5%）,其余5例经足背静脉留置针推注造影剂顺行造影后穿刺成功。经3~7 d溶栓治疗,患者溶栓前、后的小腿周径差分别为（4.19±1.51）、（1.38±0.50）cm,大腿周径差分别为（6.07±1.78）、（2.22±1.22）cm,差异均有统计学意义（t=9.21、9.28,P值均<0.001）;患者溶栓前、后患肢静脉通畅度评分分别为（7.41±1.55）、（2.04±0.85）分,差异有统计学意义（t=15.76,P<0.001）;溶栓后患肢静脉通畅度为70.62%±14.55%。溶栓前、后腘静脉通畅度评分分别为（5.04±1.01）、（1.26±0.71）分,差异有统计学意义（t=15.42,P<0.001）;溶栓后腘静脉通畅度73.21%±17.05%。患者住院溶栓期间均未出现大出血、小腿血肿、血栓进展等严重不良反应。结论 彩色多普勒超声导引下小腿胫/腓静脉-腘静脉顺行置管溶栓治疗急性混合型下肢DVT溶栓疗效好,对下肢静脉通畅度,特别是腘静脉通畅度改善明显,是一种安全有效的微创治疗方法,具有较好的临床应用价值。     

The cathepsin-S/protease-activated receptor-(PAR)-2 axis drives chronic allograft vasculopathy and is a molecular target for therapeutic intervention

BackgroundCathepsin S (CatS) and proteinase-activated receptor (PAR)-2 are involved in the remodelling of vascular walls and neointima formation as well as in alloantigen presentation and T-cell priming. Therefore, we hypothesized that CatS/PAR-2 inhibition/deficiency would attenuate chronic allograft vasculopathy.MethodsHeterotopic aortic murine transplantation was performed from C57BL/6J donors to C57BL/6J recipients (syngeneic control group), Balb/c to C57BL/6J without treatment (allogenic control group), Balb/c to C57BL/6J with twice daily oral CatS inhibitor (allogenic treatment group) and Balb/c to Par2−/− C57BL/6J (allogenic knockout group). The recipients were sacrificed on day 28 and the grafts were harvested for histological analysis and RT-qPCR.ResultsAfter 28 days, mice of the allogenic control group exhibited significant neointima formation and massive CD8 T-cell infiltration into the neointima while the syngeneic control group showed negligible allograft vasculopathy. The mRNA expression level of CatS in allografts was 5-fold of those in syngeneic grafts. Neointima formation and therefore intima/media-ratio were significantly decreased in the treatment and knockout group in comparison to the allogenic control group. Mice in treatment group also displayed significantly fewer CD8 T cells in the neointima compared with allogeneic controls. Additionally, treatment with the CatS inhibitor and PAR2-deficiency decreased mRNA-levels of interleukins and cytokines.ConclusionIn conclusion, our data indicate that inhibiting CatS and PAR-2 deficiency led to a marked reduction of neointima formation and associated inflammation in a murine heterotopic model for allograft vasculopathy.     

Let-7b-5p inhibits colon cancer progression by prohibiting APC ubiquitination degradation and the Wnt pathway by targeting NKD1

Naked cuticle homolog 1 (NKD1), which is expressed at low levels in many tumors, is considered an inhibitor of the Wnt/β-catenin pathway, but it is highly expressed in colon cancer and can promote colon cancer cell proliferation. miRNAs are involved in the occurrence and progression of many tumors. However, miRNAs that can regulate NKD1 and the mechanisms by which NKD1 regulates tumor progression remain ambiguous. This research aims to reveal the potential regulatory network of NKD1 in colon cancer. miRNA data downloaded from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases were analyzed by bioinformatics to screen for potential miRNAs targeting NKD1. Let-7b-5p was found to inhibit proliferation, migration, and invasion of colon cancer cells targeting NKD1. Further studies suggested that let-7b-5p can modulate Wnt signaling activity, and the nuclear accumulation of β-catenin was significantly restrained by let-7b-5p through targeting NKD1. Moreover, NKD1 could prohibit the expression of the APC protein. Further studies manifested that NKD1 bound to APC and promoted the ubiquitination degradation of APC through restraining the expression of the deubiquitinating enzyme USP15 and blocking the combination between USP15 and APC. Functionally, NKD1 enhanced the proliferation and migration of colon cancer cells by inhibiting APC expression. This research revealed a novel mechanism by which the let-7b-5p-NKD1-APC-β-catenin signaling pathway inhibited colon cancer cell progression.     

Comprehensive analysis of circular RNA-associated competing endogenous RNA networks and immune infiltration in gastric cancer

BackgroundCircular RNA (circRNA) has been proved to be an important regulator of gastric cancer (GC). However, the role and regulatory mechanism of circrna related competitive endogenous RNA (ceRNA) in GC have not been established.MethodsCircRNA data and clinical data were obtained from the GEO and TCGA databases. The ceRNA networks were constructed and a function enrichment analysis was completed. Additionally, correlations between hub genes expression, immune cell infiltration, and clinical phenotypes were determined. The differentially expressed circRNAs and their downstream microRNAs (miRNAs) were validated by quantitative real-time polymerase chain reaction, and the hub genes were validated by western blot analysis. The migration and invasion ability of overexpressed hsa_circ_0002504 was determined by a transwell assay.ResultsThe ceRNA network contained 2 circRNAs, 3 miRNAs, and 55 messenger RNAs (mRNAs). 323 biological processes terms, 53 cellular components terms, 51 molecular functions terms, and 4 signaling pathways were revealed by the function enrichment analysis. The GSEA analysis revealed that the hub genes were positively correlated with the axon guidance and adhesion molecules pathways. The correlation analysis revealed that overexpressed EPHA4 and KCNA1 indicated poor tissue differentiation and were associated with clinically advanced stages of GC. The in vitro experiments showed that hsa_circ_0002504 was significantly down-regulated in GC cell lines. In addition, the overexpression of hsa_circ_0002504 led to a significant downregulation of hsa-miR-615-5p and hsa-miR-767-5p, as well as an upregulation of EPHA4, KCNA1, and NCAM1. Furthermore, it suppressed the migration and invasion ability of GC cells.ConclusionsHsa_circ_0002504 is a potential diagnostic biomarker for GC. High expression of EPHA4 and KCNA1 may indicate poor prognosis.     

CD8+iTregs mediate the protective effect of rapamycin against graft versus host disease in a humanized murine model

CD8⁺Tregs are important immunoregulatory cells that participate in immunopathological processes in many diseases. Rapamycin (Rapa) is a macrolide immunosuppressant that inhibits the mammalian target of rapamycin (mTOR) and has been shown to improve CD4⁺-induced Tregs (iTregs) generation. This study aimed to evaluate the role of Rapa in the generation and function of CD8⁺iTregs. Human CD8 + CD25-CD45RA + T cells were divided into two groups, one with Rapa and the other without Rapa, and both groups were cultured under Treg-induced conditions. Rapa significantly improved Foxp3 expression and the suppressive function of CD8⁺iTregs in vitro. Further studies showed that Rapa suppressed inflammatory cytokine expression and enhanced anti-inflammatory cytokine expression. Under inflammatory conditions in vitro, Rapa-CD8 + iTregs sustained Foxp3 and anti-inflammatory cytokine expression. An in-depth study showed that Rapa regulated CpG demethylation in the Foxp3 region and STAT1 and STAT3 phosphorylation in CD8⁺iTregs. Finally, we compared the regulatory ability of Rapa and all-trans retinoic acid, another reagent that stimulates CD4⁺ iTreg generation in vitro, which showed that Rapa, but not all-trans retinoic acid, improved CD8⁺ iTreg induction and suppressed CD4⁺T cell expansion in vitro and protected against graft-versus-host disease in a humanized murine model in vivo. These results strongly suggest that CD8⁺iTregs initiated by Rapa may represent a new therapeutic strategy for inflammatory and autoimmune diseases.     

Circular RNA circ_0114876 regulates osteoarthritis through upregulating ADAM10 via targeting miR-1227-3p

BackgroundOsteoarthritis (OA) was a chronic degenerative joint disease. The dysregulation of circular RNAs (circRNAs) has been identified in OA progression. However, the function and regulation mechanism of circ_0114876 in OA remains largely unknown.MethodFirstly, we used LPS-treated C28/I2 cells as a cellular model of OA. Quantificational real-time polymerase chain reaction (qRT-PCR) was used to determine the expression levels of circ_0114876, miRNA-1227-3p, and ADAM10 in OA chondrocytes. Cell Counting Kit-8 (CCK8), 5-ethynyl-20-deoxyuridine (EdU) incorporation assays, flow cytometry, Enzyme-linked immunosorbent assay (ELISA) kit, and western blot were applied to confirm cell proliferation, apoptosis, inflammation, and extracellular matrix.of circ_0114876 in vitro. The interaction between circ_0114876 and its downstream target (miR-1227-3p) and mRNA target ADAM metallopeptidase domain 10 (ADAM10), was evaluated by luciferase assay and RNA immunoprecipitation (RIP) assay.ResultCirc_0114876 and ADAM10 were upregulated and miR-1227-3p was decreased in OA tissues and LPS-treated chondrocytes. Low expression of circ_0114876 promoted proliferation and inhibited apoptosis, inflammation, and extracellular matrix of the LPS-treated chondrocytes. Mechanistically, circ_0114876 functioned in human chondrocytes through targeting miR-1227-3p and ADAM10. Furthermore, miRNA-1227-3p inhibitor reversed the effect of circ_0114876 knockdown on the OA chondrocytes, and ADAM10 overexpression reversed the effect of miR-1227-3p mimic on the OA chondrocytes.ConclusionCirc_0114876 was increased in OA tissues and cells. Circ_0114876 facilitated the progression in the LPS-induced OA cell model via regulating the miR-1227-3p/ADAM10 axis. This study would provide a potentially effective therapeutic strategy for OA progression.     

Effect of blood transfusion post kidney transplantation on de novo human leukocytes antigen donor-specific antibody development and clinical outcomes in kidney transplant recipients: A systematic review and meta-analysis

The relationship between blood transfusion following kidney transplantation (KT) and the development of de novo donor-specific antibodies (dnDSA) is controversial. This was investigated by conducting a meta-analysis of studies on patients who underwent KT with or without blood transfusion, and by evaluating the effect of post-KT blood transfusion on clinical outcomes of kidney transplant recipients. Relevant studies in the PubMed, EMBASE, and Cochrane Library databases were identified from inception to July 1, 2022. Two reviewers independently extracted data from the selected articles and estimated study quality. A fixed effects or random effects model was used to pool data according to the heterogeneity among studies. Data included in the meta-analysis were derived from 11 studies with a total of 19,543 patients including 6191 with and 13,352 without blood transfusion post-KT. We assessed the pooled associations between blood transfusion and occurrence of dnDSA and clinical outcomes of transplant recipients. Blood transfusion was strongly correlated with the development of dnDSA (relative risk [RR] = 1.40, 95% confidence interval [CI]: 1.17–1.67; P < 0.05). Patients with blood transfusion had a higher risk of developing anti-human leukocyte antigen (HLA) class I dnDSA than non-transfused patients (RR = 1.75, 95% CI: 1.14–2.69; P < 0.05) as well as significantly higher rates of antibody-mediated rejection (AMR) (RR = 1.41, 95% CI: 1.21–2.35; P < 0.05) and graft loss (RR = 1.75, 95% CI: 1.30–2.35; P < 0.05). There were no statistically significant differences between the two groups in the development of anti-HLA antibodies, anti-HLA class II dnDSA, and anti-HLA class I and II dnDSA; delayed graft function; T cell-mediated rejection; acute rejection; borderline rejection; or patient death. Our results suggest that blood transfusion was associated with dnDSA development in KT recipients. The findings of this systematic review also suggest that post-KT blood transfusion recipients have a higher risk of AMR, and graft loss compared with non-transfused patients. Evidence from this meta-analysis indicates that the use of blood transfusion post-KT is associated with a significantly higher risk of immunological sensitization. More and higher quality results from large randomized controlled trials are still needed to inform clinical practice.     

Rejection and graft outcomes in kidney transplant recipients with and without angiotensin II receptor type 1 antibodies

AimAngiotensin II type 1 receptor antibody (AT1R Ab) is a non-Human Leucocyte Antigen (HLA) antibody that is maybe associated with early severe kidney transplant rejection and worse graft outcomes. This study aimed to assess the association between AT1R Ab and kidney transplant rejection and graft outcomes.MethodsWe performed a retrospective analysis of all adult kidney transplant recipients in an Australian centre who had an AT1R Ab test between 1 January 2015 to 30 June 2020. AT1R Ab positive patients were compared to AT1R Ab negative patients. Primary outcomes were rejection risk, type and histopathological severity scores. Secondary outcomes were 8-week graft function and graft loss.ResultsOf 965 kidney transplants that were performed during the study period, 73 patients had AT1R Ab tested; 16 (22%) were positive and 57(78%) were negative. Positive patients were on average younger and had higher level of donor-specific HLA antibodies. Rejection occurred in 13 (81%) positive patients and 41 (72%) negative patients (P = 0.45). No significant differences in rejection type or severity were found. HLA mismatch and peak panel reactive antibody ≥80%, but not AT1R Ab, independently predicted rejection. Average (132 vs. 177 mmol/L, P = 0.302) and graft loss were not significantly different between groups.ConclusionThe study found no evidence that AT1R Ab is associated with rejection type, severity or worse graft function. Future studies should assess its relationship with graft outcomes to help complement immunological risk assessment and potentially provide therapeutic options to alter outcomes.     

mCTA侧支循环评分及血清microRNA-134、VEGF、bFGF水平预测AIS大脑中动脉闭塞患者预后的价值

目的 探究多时相计算机断层扫描血管成像（mCTA）侧支循环评分及血清microRNA-134（miR-134）、血管内皮细胞生长因子（VEGF）和碱性成纤维细胞生长因子（bFGF）水平预测急性缺血性脑卒中（AIS）大脑中动脉闭塞患者预后的价值。方法 选取2020年2月—2023年2月在江苏大学附属武进医院住院治疗的AIS大脑中动脉闭塞患者108例。检测患者治疗期间的mCTA侧支循环评分及血清miR-134、VEGF、bFGF水平，并进行随访。根据患者出院后3个月的改良Rankin量表评分，分为预后良好组（改良Rankin量表评分≤ 2分，47例）、预后不良组（改良Rankin量表评分> 2分，61例），对可能影响患者预后的因素进行分析，并绘制ROC曲线分析其诊断价值。结果 预后不良组最终梗死体积大于预后良好组（P <0.05），mCTA侧支循环评分低于预后良好组（P <0.05）；预后不良组miR-134相对表达量高于预后良好组（P <0.05），VEGF、bFGF水平均低于预后良好组（P <0.05）。预后不良组年龄、低密度脂蛋白水平高于预后良好组（P <0.05）。多因素一般Logistic回归分析结果显示：mCTA侧支循环评分［O^R=0.804（95% CI：0.729，0.974）］、VEGF［O^R=0.618（95% CI：0.397，0.963）］、bFGF［O^R=0.608（95% CI：0.402，0.919）］为AIS大脑中动脉闭塞患者预后良好的保护性因素（P <0.05）；miR-134［O^R=1.941（95% CI：1.802，3.480）］、低密度脂蛋白［O^R=1.349（95% CI：1.051，1.730）］是AIS大脑中动脉闭塞患者预后不良的危险因素（P <0.05）。ROC曲线分析结果表明，mCTA侧支循环评分、miR-134、VEGF、bFGF预测AIS大脑中动脉闭塞患者预后不良的曲线下面积分别为0.843、0.946、0.937和0.892，敏感性分别为7.66%（95% CI：0.695，0.837）、9.36%（95% CI：0.900，0.972）、8.72%（95% CI：0.823，0.921）、7.23%（95% CI：0.661，0.785），特异性分别为83.6%（95% CI：0.770，0.902）、82.0%（95% CI：0.770，0.870）、86.9%（95% CI：0.818，0.920）、93.4%（95% CI：0.896，0.972）。结论 预后不良患者最终梗死体积较大，mCTA侧支循环评分较低，血清miR-134、VEGF和bFGF水平较低。mCTA侧支循环评分、血清miR-134、VEGF、bFGF水平对AIS大脑中动脉闭塞患者预后不良有较好的预测价值，可作为预后评估的指标。