外周血循环肿瘤DNA检测方法及应用

肿瘤是一种威胁人类健康的重大疾病。研究发现,肿瘤的发生和发展是基于分子水平上癌基因(oncogenes)的激活和抑癌基因(anti-oncogene)的失活所致。基因突变分析对于肿瘤诊断和预后至关重要。早期诊断被认为是提高癌症病人治愈率,降低其死亡率最有效的途径。外周血循环肿瘤核酸中可检测到与肿瘤细胞一致的基因突变。这些基因突变的检测可以为肿瘤的分期及药物的治疗疗效和预后提供丰富的信息。因此,外周血循环肿瘤DNA突变检测为肿瘤的早期诊断和治疗提供了新的研究方向和领域。     

The efficacy and safety of Batroxobin in combination with anticoagulation on cerebral venous sinus thrombosis

Cerebral venous sinus thrombosis (CVST) is an uncommon subtype of stroke with highly variable clinical presentation. Although anticoagulation with heparin and/or warfarin remains the standard treatment for CVST, treatment failure is still common. This study aims to evaluate the safety and efficacy of Batroxobin in combination with anticoagulation on CVST control. In this retrospective study, a total of 61 CVST patients were enrolled and divided into Batroxobin (n?=?23) and control (n?=?38) groups. In addition to the same standard anticoagulation in control, patients in the treatment group received Batroxobin 5 BU intravenous infusion (10 BU for the first time) every other day, for a total of three infusions. A higher recanalization rate was found in Batroxobin group (adjusted OR [95% CI] of 2.5 [1.1–5.0], p?=?0.028) compared to the control group, especially in patients with high levels of fibrinogen (adjusted OR [95% CI] of 4.7 [1.4–16.7], p?=?0.015). Statistically significant differences between the two groups were seen regarding the levels of thrombin time, fibrinogen and d-dimer at each cut-off time point (all p?<?0.01). Compared with baseline, NIHSS scores at discharge showed significant improvement in the Batroxobin group [0(0, 4.25)–5(2, 11), p?=?0.036]. No significant difference in mRS scores was found between the two groups at discharge or at 6-month outpatient follow-up (all p?>?0.05). Additionally, Batroxobin did not increase the risk of intracranial hemorrhage. We conclude that Batroxobin is a potentially safe and effective adjunct therapeutic agent promoting CVST recanalization especially in patients with high level of fibrinogen.     

永存原始三叉动脉供血颈动脉-海绵窦瘘1例北大核心CSCD

1临床资料患者女,51岁。因“左侧头部及面部搏动性疼痛伴头晕、耳鸣2个月”入院。患者既往身体健康,查体可闻及左侧眼球后搏动性血管杂音,左侧眼球突出伴球结膜水肿。患者入院后行全脑血管造影见左侧永存原始三叉动脉供血的颈动脉-海绵窦瘘(carotid-cavernous sinus fistula,CCF),原始三叉动脉两端分别与左侧颈内动脉海绵窦段及左侧小脑上动脉相连,左侧小脑上动脉远端未见显影,主要经眼上静脉、岩上窦、基底窦、直窦引流(图1)。     

Development of novel quality control material based on CRISPR/Cas9 editing and xenografts for MLH1 protein deficiency testing

BackgroundMismatch repair deficiency (dMMR) status induced by MLH1 protein deficiency plays a pivotal role in therapeutic decision‐making for cancer patients. Appropriate quality control (QC) materials are necessary for monitoring the accuracy of MLH1 protein deficiency assays used in clinical laboratories.MethodsCRISPR/Cas9 technology was used to edit the MLH1 gene of GM12878Cas9 cells to establish MLH1 protein‐deficient cell lines. The positive cell lines were screened and validated by Sanger sequencing, Western blot (WB), and next‐generation sequencing (NGS) and were then used to prepare formalin‐fixed, paraffin‐embedded (FFPE) samples through xenografting. These FFPE samples were tested by hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) for suitability as novel QC materials for MLH1 protein deficiency testing.ResultsWe successfully cultured 358 monoclonal cells, with a survival rate of 37.3% (358/960) of the sorted monoclonal cells. Through Sanger sequencing, cell lines with MLH1 gene mutation were identified. Subsequently, two cell lines with MLH1 protein deficiency were identified by WB and named as GM12878Cas9_6 and GM12878Cas9_10. The NGS results further confirmed that the MLH1 gene mutation in these two cell lines would cause the formation of stop codons and terminate the expression of the MLH1 protein. The H&E staining and IHC results also verified the deficiency of the MLH1 protein, and FFPE samples from xenografts proved their similarity and consistency with clinical samples.ConclusionsWe successfully established MLH1 protein‐deficient cell lines. Followed by xenografting, we developed novel FFPE QC materials with homogenous, sustainable, and typical histological structures advantages that are suitable for the standardization of clinical IHC methods.     

Recurrent high fever with macular papules in an elderly male: a case report

Immunologic Research -     

Allogeneic Vγ9Vδ2 T-cell immunotherapy exhibits promising clinical safety and prolongs the survival of patients with late-stage lung or liver cancer

Vγ9Vδ2 T cells are promising candidates for cellular tumor immunotherapy. Due to their HLA-independent mode of action, allogeneic Vγ9Vδ2 T cells can be considered for clinical application. To apply allogeneic Vγ9Vδ2 T cells in adoptive immunotherapy, the methodology used to obtain adequate cell numbers with optimal effector function in vitro needs to be optimized, and clinical safety and efficacy also need to be proven. Therefore, we developed a novel formula to improve the expansion of peripheral γδ T cells from healthy donors. Then, we used a humanized mouse model to validate the therapeutic efficacy of expanded γδ T cells in vivo; furthermore, the expanded γδ T cells were adoptively transferred into late-stage liver and lung cancer patients. We found that the expanded cells possessed significantly improved immune effector functions, including proliferation, differentiation, and cancer cell killing, both in vitro and in the humanized mouse model. Furthermore, a phase I clinical trial in 132 late-stage cancer patients with a total of 414 cell infusions unequivocally validated the clinical safety of allogeneic Vγ9Vδ2 T cells. Among these 132 patients, 8 liver cancer patients and 10 lung cancer patients who received ≥5 cell infusions showed greatly prolonged survival, which preliminarily verified the efficacy of allogeneic Vγ9Vδ2 T-cell therapy. Our clinical studies underscore the safety and efficacy of allogeneic Vγ9Vδ2 T-cell immunotherapy, which will inspire further clinical investigations and eventually benefit cancer patients.     

细胞外囊泡在组织工程研究中的新应用

细胞外囊泡(EVs)是细胞分泌释放的微小囊泡,其中包含核酸、蛋白质等生物活性分子,在细胞间通讯和信号传导中发挥着重要的作用。EVs可以通过递送生物活性分子,在多种疾病中起到治疗作用,有望成为新一代药物递送的天然载体。相较于传统纳米材料,天然EVs靶向递送具有低免疫原性、良好生物相容性、天然靶向性、长循环时间等明显优势。通过进一步组织工程手段来个性化构建EVs,提高其靶向性和目标治疗分子负载能力,为靶向分子治疗提供了一种新的治疗策略。