Prognostic Value of Disseminated Tumor Cells in the Bone Marrow of Patients with Operable Primary Breast Cancer: A Long-term Follow-up Study

Background

Detection of disseminated tumor cells (DTC) in primary breast cancer (BC) patients’ bone marrow (BM) seems to be a surrogate marker of tumor spread and an independent prognostic factor for disease-free and overall survival.

Methods

Here we present the largest single-center cohort of patients (n = 1378) with the longest observation time (median 82.0 months). Immunocytochemical staining was performed using murine monoclonal antibody 2E11 with the avidin–biotin complex technique.

Results

At primary surgery, 49 % of patients showed MUC-1 positive cells inside their BM. Patients without BM DTC had significantly more often T1-tumors (P = 0.007) with less often affected axillary lymph nodes (P < 0.001). We observed a significantly higher incidence of distant metastases in DTC positive patients (P < 0.001). This leads to a reduced disease-free survival (P < 0.0001). Furthermore, in DTC positive patients there was a higher mortality rate and, accordingly, a reduced overall survival (P < 0.0001).

Conclusions

Due to the presence of BM DTC, patients with a clinically poorer outcome can be identified at primary surgery. We therefore suggest that DTC analysis can be used as a prognostic factor and monitoring tool in clinical trials. Future study concepts relating to DTC should aim at identification of BC patients who may profit from adjuvant systemic therapy.     

Bone Regeneration Based on Tissue Engineering Conceptions — A 21st Century Perspective

The role of Bone Tissue Engineering in the field of Regenerative Medicine has been the topic of substantial research over the past two decades. Technological advances have improved orthopaedic implants and surgical techniques for bone reconstruction. However, improvements in surgical techniques to reconstruct bone have been limited by the paucity of autologous materials available and donor site morbidity. Recent advances in the development of biomaterials have provided attractive alternatives to bone grafting expanding the surgical options for restoring the form and function of injured bone. Specifically, novel bioactive (second generation) biomaterials have been developed that are characterised by controlled action and reaction to the host tissue environment, whilst exhibiting controlled chemical breakdown and resorption with an ultimate replacement by regenerating tissue. Future generations of biomaterials (third generation) are designed to be not only osteoconductive but also osteoinductive, i.e. to stimulate regeneration of host tissues by combining tissue engineering and in situ tissue regeneration methods with a focus on novel applications. These techniques will lead to novel possibilities for tissue regeneration and repair. At present, tissue engineered constructs that may find future use as bone grafts for complex skeletal defects, whether from post-traumatic, degenerative, neoplastic or congenital/developmental “origin” require osseous reconstruction to ensure structural and functional integrity. Engineering functional bone using combinations of cells, scaffolds and bioactive factors is a promising strategy and a particular feature for future development in the area of hybrid materials which are able to exhibit suitable biomimetic and mechanical properties. This review will discuss the state of the art in this field and what we can expect from future generations of bone regeneration concepts.     

Accumulation of methotrexate polyglutamates in lymphoblasts is a determinant of antileukemic effects in vivo. A rationale for high-dose methotrexate.

Methotrexate (MTX) is one of the most widely used drugs for the treatment of childhood acute lymphoblastic leukemia (ALL) and is commonly given in high doses. However, the rationale for high-dose MTX (HDMTX) has been challenged recently. To determine whether higher MTX polyglutamate (MTXPG) concentrations in ALL blasts translate into greater antileukemic effects, 150 children with newly diagnosed ALL were randomized to initial treatment with either HDMTX (1,000 mg/m2 intravenously over 24 h) or lower-dose MTX (30 mg/m2 by mouth every 6 h x 6). ALL blasts accumulated higher concentrations of MTXPG and long-chain MTXPG (MTXPGLC) after HDMTX (P < 0.00001). Of 101 patients evaluable for peripheral blast cytoreduction, MTXPG concentrations were higher in patients whose blast count decreased within 24 h (P = 0.005) and in those who had no detectable circulating blasts within 4 days (P = 0.004). The extent of inhibition of de novo purine synthesis in ALL blasts was significantly related to the blast concentration of MTXPGLC (IC95% = 483 pmol/10(9) blasts). The percentage of patients with 44-h MTXPGLC exceeding the IC95% was greater after HDMTX (81%) than LDMTX (46%, P < 0.0001). These data indicate that higher blast concentrations of MTXPG are associated with greater antileukemic effects, establishing a strong rationale for HD-MTX in the treatment of childhood ALL.     

Protein-tyrosine kinase p72syk in Fc gamma RI receptor signaling

In this report we show that gamma-interferon (IFN) induces the expression of the nonreceptor protein tyrosine kinase, p72syk, and that cross-linking the Fc gamma RI receptor in IFN-differentiated U937 cells (U937IF cells) results in the activation of syk kinase. We show that syk is tyrosine phosphorylated (12-fold increase) after Fc gamma RI cross-linking. In vitro kinase assays demonstrate that the specific kinase activity of syk increased eightfold after Fc gamma RI cross- linking. The activation of signal transduction through the Fc gamma RI receptor, as measured by the respiratory burst, is associated with the tyrosine phosphorylation and catalytic activation of the syk kinase. We show that syk coprecipitates with the gamma subunit of the Fc gamma RI, Fc gamma RI gamma. The data suggest that p72syk is involved in signal transduction through the Fc gamma RI receptor, involving the Fc gamma RI gamma subunit.     

Endocarditis Caused by Rhodotorula Infection

Rhodotorula is an emerging opportunistic fungal pathogen that is rarely reported to cause endocarditis. We describe a case involving a patient who developed endocarditis due to Rhodotorula mucilaginosa and Staphylococcus epidermidis, proven by culture and histopathology. The case illustrates the unique diagnostic and therapeutic challenges relevant to Rhodotorula spp.     

Functional characterization of novel rare CYP2A6 variants and potential implications for clinical outcomes

CYP2A6 activity, phenotyped by the nicotine metabolite ratio (NMR), is a predictor of several smoking behaviors, including cessation and smoking‐related disease risk. The heritability of the NMR is 60–80%, yet weighted genetic risk scores (wGRSs) based on common variants explain only 30–35%. Rare variants (minor allele frequency <1%) are hypothesized to explain some of this missing heritability. We present two targeted sequencing studies where rare protein‐coding variants are functionally characterized in vivo, in silico, and in vitro to examine this hypothesis. In a smoking cessation trial, 1687 individuals were sequenced; characterization measures included the in vivo NMR, in vitro protein expression, and metabolic activity measured from recombinant proteins. In a human liver bank, 312 human liver samples were sequenced; measures included RNA expression, protein expression, and metabolic activity from extracted liver tissue. In total, 38 of 47 rare coding variants identified were novel; characterizations ranged from gain‐of‐function to loss‐of‐function. On a population level, the portion of NMR variation explained by the rare coding variants was small (~1%). However, upon incorporation, the accuracy of the wGRS was improved for individuals with rare protein‐coding variants (i.e., the residuals were reduced), and approximately one‐third of these individuals (12/39) were re‐assigned from normal to slow metabolizer status. Rare coding variants can alter an individual’s CYP2A6 activity; their integration into wGRSs through precise functional characterization is necessary to accurately assess clinical outcomes and achieve precision medicine for all. Investigation into noncoding variants is warranted to further explain the missing heritability in the NMR.

Study Highlights

• WHAT IS THE CURRENT KNOWLEDGE ON THE TOPIC?

Common CYP2A6 variants (minor allele frequency >1%) explain 30–35% of the variation in CYP2A6 activity, despite high heritability estimates (60–80%) in the CYP2A6 activity biomarker measure. One hypothesis is that rare coding variants (minor allele frequency <1%) may explain a portion of the missing heritability from pharmacogenes, including CYP2A6.

• WHAT QUESTION DID THIS STUDY ADDRESS?

What is the relative contribution of rare coding variants in explaining variation in CYP2A6 activity? How necessary is the incorporation of rare coding variants in predicting individual metabolic status, and consequent tailoring of treatment?

• WHAT DOES THIS STUDY ADD TO OUR KNOWLEDGE?

Rare coding variants may explain only a small fraction of the variation on a population level; however, their role may be important on an individual level, altering the predicted metabolic status in a third of the individuals with these rare coding variants.

• HOW MIGHT THIS CHANGE CLINICAL PHARMACOLOGY OR TRANSLATIONAL SCIENCE?

Evaluating rare coding variants in pharmacogenes, such as CYP2A6, will be valuable in enhancing the investigation of CYP2A6’s influence on tobacco addiction and disease pathogenesis, by providing a more accurate reflection of the phenotypic metabolic status through improved genetic assessments.     

不同载荷作用下C（4～5）椎间孔孔径的变化

目的:分析不同载荷对C4～5椎间孔孔径变化的影响。方法:实验于2006-01/2006-05在湘南学院附属医院进行。成人新鲜尸体颈椎标本10具(自愿捐献或家属同意),节段包括C3～T1,剔除肌肉组织,保留椎间盘、韧带和关节囊结构的完整。分别测量椎间盘完整、椎间盘髓核摘除、颈人工椎间盘置换和前路钢板植骨内固定4种状态下,加压速度为5mm/min时,25,50,75,100,125和150N的分级轴向载荷加载于标本时C4～5上下径、上前后径、下前后径以及椎间孔面积的变化。结果:①轴向加载25N时,C4～5椎间孔面积椎间盘髓核摘除标本小于完整椎间盘、颈人工椎间盘置换和前路钢板植骨内固定标本[(57.26±17.65),(65.81±16.83),(75.37±17.75),(66.21±16.50)mm2,P<0.05];颈人工椎间盘置换标本大于颈椎植骨融合钢板内固定标本(P<0.05)。②轴向加载50N时,C4～5椎间孔面积椎间盘髓核摘除标本小于其他3组[(57.13±17.64),(65.72±16.85),(75.35±17.75),(66.11±16.46)mm2,P<0.05];颈人工椎间盘置换标本大于颈椎植骨融合钢板内固定标本(P<0.05)。③轴向加载75N时,上下径:C4～5椎间孔面积椎间盘髓核摘除标本小于其他3组[(55.26±17.66),(64.80±16.85),(73.32±17.74),(65.21±16.48)mm2,P<0.05];颈人工椎间盘置换标本大于颈椎植骨融合钢板内固定标本(P<0.05)。④轴向加载100N时,上下径:C4～5椎间孔面积椎间盘髓核摘除标本小于其他3组[(53.22±17.66),(63.81±16.83),(71.35±17.76),(65.27±16.46)mm2,P<0.05]。⑤轴向加载125N时,C4～5椎间孔面积椎间盘髓核摘除标本小于其他3组[(51.25±17.64),(62.82±16.83),(69.25±17.74),(65.25±16.43)mm2,P<0.05]。⑥椎间孔面积:轴向加载150N时,C4～5椎间孔面积椎间盘髓核摘除标本小于其他3组[(49.16±17.65),(61.84±16.86),(67.15±17.73),(65.24±16.42)mm2,P<0.05]结论:颈椎间盘髓核摘除后C4～5椎间孔有效空间明显减少。     

自体骨髓单个核细胞经介入途径移植治疗股骨头坏死54例：12个月疗效随访

目的:观察经介入途径移植自体骨髓单个核细胞在股骨头坏死治疗中的应用,并评价其疗效。方法:选择2004-07/2005-11在解放军四六三院细胞治疗中心住院的,具有完整随访资料的股骨头坏死确诊患者共54例91髋。纳入确诊股骨头坏死,有关节疼痛、功能障碍等症状患者,性别、年龄不限;排除有严重心力衰竭、严重肾功能异常等不能耐受手术者。符合纳入标准54例,男45例,女9例,12～68岁。按ARCO分期Ⅱ期42髋,Ⅲ期47髋,Ⅳ期2髋。实验对象对治疗的相关内容知情同意并签知情同意。干预措施:抽取患者髂后上嵴骨髓进行单个核细胞悬液的制备。在DSA监视下将采集的单个核细胞混悬液经股动脉行Seldinger法穿刺,穿刺成功后,置入4F动脉鞘,经动脉鞘置入Cobra导管,将导管超选择至闭孔动脉及旋股内外侧动脉,平均注入单个核细胞悬液。术后定期随访症状变化情况,1年后复查X射线或CT,随访疼痛、关节活动度等情况。实验评估:①疼痛指数:无疼痛症状为3分,Harris髋关节评分疼痛分级A级;时有隐痛2分,Harris髋关节评分疼痛B级;轻度疼痛为1分,Harris髋关节评分疼痛C级;中度疼痛为0分,Harris髋关节评分疼痛D级。②功能指数:髋关节屈、伸、展、收、旋转度评分达Harris髋关节活动范围评分4～5分为3分;3～4分为2分;2～3分为1分;小于2分为0分。③X射线平片指数:股骨头形态无变化,应力骨小梁清晰,坏死区明显缩小为3分;坏死区略缩小为2分;治疗前后无明显变化为1分;坏死区扩大为0分。④血管指数:治疗后旋股内、外侧动脉及其分支增粗、增多,延长1cm以上者3分;1～0.5cm者2分;小于0.5cm者1分,无变化者0分。结果:54例患者均完成疼痛症状、关节功能及影像学随访1年。①术后12个月复查疼痛消失9髋,缓解61髋,无缓解21髋,缓解率为76.9%。②关节功能缓解33髋,无缓解58髋,缓解率为36.3%。③1年后X射线平片或CT、MRI示股骨头区可见不同程度的股骨头坏死区骨质密度改变,坏死区有吸收、缩小,股骨头形态变圆滑规整,改善28髋,无缓解或加重63髋,缓解率为30.1%。④12例24髋完成术后12个月复查股骨头供血动脉数字减影血管造影,好转18髋,好转率为72.2%。结论:经介入途径移植自体骨髓单个核细胞治疗股骨头坏死损伤小,可缓解临床相关症状。