Analysis of vascularization in thyroid gland nodes with superb microvascular imaging (SMI) and CD34 expression histology: a pilot study

Background The Doppler sonography technique known as "superb microvascular imaging" (SMI) is advancing sonographic micro vascularization imaging in various disciplines. In this study, we aimed to determine whether SMI could reliably reproduce the blood flow in thyroid nodes and whether malignancy could be diagnosed, based on vascularization properties. Immunhistochemical staining by CD34 and SMI where used to determine the vascularization of nodes in terms of quantified vascularization parameters gained by computational evaluation. Methods We used image analysis programs to investigate whether the quantitative value for vascularization strength in the thyroid node, measured with SMI, was correlated with the actual degree of vascularization, determined microscopically. We included 16 patients that underwent thyroid resections. We prepared thyroid gland tissue slices for immunohistochemistry and labelled endothelial cells with CD34 to visualize blood vessels microscopically. We used image analysis programs, ImageJ, to quantify SMI Doppler sonographic measurements and CellProfiler to quantify CD34 expression in histological sections. We evaluated the numeric values for diagnostic value in node differentiation. Furthermore, we compared these values to check for correlations. Results Among the 16 nodes studied, three harboured malignant tumours (18.75%): two papillary and one follicular carcinoma. Among the 13 benign lesions (81.25%), four harboured follicular adenomas. Malignant and benign nodes were not significantly different in sonographic (0.88 ± 0.89 vs. 1.13 ± 0.19; p = 0.2790) or immunohistochemical measurements of vascularization strength (0.05 ± 0.05 vs. 0.08 ± 0.06; p = 0.2260). Conclusion We found a positive, significant correlation (r = 0.55588; p = 0.0254) between SMI (quantitative values for vascularization strength) and immunohistochemistry (CD34 staining) evaluations of thyroid nodes.

Polymorphous Low-Grade Neuroepithelial Tumor of the Young (PLNTY): Molecular Profiling Confirms Frequent MAPK Pathway Activation

Polymorphous low-grade neuroepithelial tumor of the young (PLNTY) is a recently described epileptogenic tumor characterized by oligodendroglioma-like components, aberrant CD34 expression, and frequent mitogen-activated protein kinase (MAPK) pathway activation. We molecularly profiled 13 cases with diagnostic histopathological features of PLNTY (10 female; median age, 16 years; range, 5–52). Patients frequently presented with seizures (9 of 12 with available history) and temporal lobe tumors (9 of 13). MAPK pathway activating alterations were identified in all 13 cases. Fusions were present in the 7 youngest patients: FGFR2-CTNNA3 (n = 2), FGFR2-KIAA1598 (FGFR2-SHTN1) (n = 1), FGFR2-INA (n = 1), FGFR2-MPRIP (n = 1), QKI-NTRK2 (n = 1), and KIAA1549-BRAF (n = 1). BRAF V600E mutation was present in 6 patients (17 years or older). Two fusion-positive cases additionally harbored TP53/RB1 abnormalities suggesting biallelic inactivation. Copy number changes predominantly involving whole chromosomes were observed in all 10 evaluated cases, with losses of chromosome 10q occurring with FGFR2-KIAA1598 (SHTN1)/CTNNA3 fusions. The KIAA1549-BRAF and QKI-NTRK2 fusions were associated respectively with a 7q34 deletion and 9q21 duplication. This study shows that despite its name, PLNTY also occurs in older adults, who frequently show BRAF V600E mutation. It also expands the spectrum of the MAPK pathway activating alterations associated with PLNTY and demonstrates recurrent chromosomal copy number changes consistent with chromosomal instability.

Prognostic value of CD34 expression status in patients with myxofibrosarcomas and undifferentiated pleomorphic sarcomas

It is controversial whether patients with myxofibrosarcomas (MFSs) have better prognoses than those with undifferentiated pleomorphic sarcomas (UPSs). No useful prognostic factors have been established to date. We therefore aimed to evaluate the prognostic value of CD34 expression status in 192 patients with MFSs and UPSs. Using the log-rank test, we showed that patients with MFSs had a significantly better overall survival than did those with UPSs when defining the former as having a > 10% myxoid component (p = 0.03), but not when defining it as having a > 50% myxoid component (p = 0.1). Under the definition of MFSs as > 10% myxoid component, the log-rank test revealed that the diagnosis of the UPS and the CD34 loss (p < 0.001) were significant adverse predictors of overall survival. As per the Cox model, the CD34 loss remained an independent prognostic factor (hazard ratio = 3.327; 95% confidence interval 1.334–8.295), while the diagnosis of the UPS was a nonsignificant confounding factor (hazard ratio = 1.084; 95% confidence interval 0.679–1.727). In conclusion, CD34 expression status is a useful prognostic factor in patients with MFS and UPS, and it should be incorporated into grading systems that are used to predict outcomes.

Comparison of in Vitro Vasculogenesis Potential between ASCs with BM-MSCs

Tube formation assay is the most widely used method as a vasculogenesis/ angiogenesis test in vitro. Mesenchymal stem cells (MSCs) are multipotent adult cells. The paracrine effect of MSCs on neovascularization is well known. In general, MSCs do not express CD34 hematopoietic surface marker, but according to some experts, bone marrow mesenchymal stem cells (BM-MSCs) express CD34 in vivo and lose their expression when they are cultured in vitro, while adipose-derived stem cells (ASCs) still have CD34 expression in the early passages when cultured in vitro. BM-MSCs are the most widely used MSC, but ASCs are also used in stem cell therapy and tissue engineering for angiogenesis purposes. Until now, the potential of vasculogenesis between ASCs and BM-MSCs is still unclear. Expression of CD34 is also unknown whether affecting the quality of tube formation. This study wanted to compare the potential of vasculogenesis between ASC and BM-MSCs through tube formation test and CD34 expression.Measurements of vasculogenesis quality showed higher tube length, number of loopsand mean number of branch points on BM-MSC. Both BM-MSCs and ASCs showed low CD34 levels.BM-MSCs showed better tube formation ability compared with ASCs. No association was found between CD34 levels and MSC vasculogenesis capability. Key words: ASCs, BM-MSCs, CD34, matrigel, tube formation.

CD34 EXPRESSION AS A SURROGATE MARKER FOR DIAGNOSIS OF INVASIVE DUCTAL CARCINOMA OF BREAST.

Background : The normal stroma of breast has enormous numbers of CD34+ brocytes therefore loss of CD34 expression in mesenchymal cells has been considered as signicant in malignant transformation and as such thought to be conclusive proof in IDC. This hypothesis leads to the identication of a new marker to diagnose cases of IDC. This study is aimed to establish the role of CD34 in diagnosis of IDC of breast. AIM: To establish the CD34 stromal expression as signicant immunohistochemical marker and to establish its efcacy as diagnostic tool to diagnose IDC. Result: A total of 50 cases of already diagnosed invasive ductal carcinoma of breast were selected and CD34 staining was performed on parafn section of all specimens. Most of the patients were in age group 35-50 years who presented with breast lumps. 50 cases of invasive ductal carcinoma were seen with 22 cases shows Lymphovascular and perineural involvement and 26 cases showed nipple areola involvement and 38 showed axillary lymph node metastasis. On histological grading of 50 malignant cases 06 cases were grade - III, 26 cases were grade- II and 18 were grade - I. Normal breast tissue was taken as internal control which show diffuse CD34 expression in stroma while there is loss of expression of CD34 in stroma in inltrating ductal carcinoma. Thereby conrms the loss of cell molecules adhesive property and signal transduction. Conclusion: There is obvious absence of expression of CD34 in malignant breast lesions. Therefore in this study we conclude that CD34 has potential to be used as surrogate diagnostic marker and management of IDC.

Difference Expressions CD34 in Acute Myeloid Leukemia Cell Culture in the Administration of Cytarabine-Daunorubicine Dose Standards

The cure rate for patients with Acute Myeloid Leukemia (AML) is 20-75%. Standard-dose cytarabine + (SDAC)-daunorubicine gives a remission rate of ± 60%, and the case of relapse is frequently found. In-vivo CD34 expression is a reliable and straightforward test that must evaluate AML patients' response to predict the response of chemotherapy + induction phase accurately. Differences in in-vitro CD34 expression are expected to be able to predict chemosensitivity in AML patients. An experimental post-test-only control group study was conducted from May to December 2019, and 8 AML subjects were found. Peripheral Blood Mononuclear Cells (PBMC) were isolated from peripheral blood samples of patients with AML collected in EDTA tubes. The PBMC isolated from peripheral blood were divided into two groups, and each group contained 106 PBMC cells in culture media. The control group (without treatment) and the SDAC-daunorubicine group were 0 + incubated for 4 hours at 37 C with a 5% CO2 atmosphere. The expression of CD34 was measured using FACSCalibur™, while + CD34+ percentage was calculated with CellQuest™ software. The percentage of CD34 in the control, SDAC + DNR, showed a significant difference with p < 0.001. This study showed a significant difference between the control group and the group + administered with the standard dose of cytarabine-daunorubicine with p < 0.001. The average CD34 expression in the + SDAC-DNR treatment group was higher than in the control group. CD34 markers cannot be used as predictors of chemosensitivity in the administration of chemotherapy.

CXCL12 enhances pregnancy outcome via improvement of endometrial receptivity in mice

Successful pregnancy inevitably depends on the implantation of a competent embryo into a receptive endometrium. Although many substances have been suggested to improve the rate of embryo implantation targeting enhancement of endometrial receptivity, currently there rarely are effective evidence-based treatments to prevent or cure this condition. Here we strongly suggest minimally-invasive intra-uterine administration of embryo-secreted chemokine CXCL12 as an effective therapeutic intervention. Chemokine CXCL12 derived from pre- and peri-implanting embryos significantly enhances the rates of embryo attachment and promoted endothelial vessel formation and sprouting in vitro. Consistently, intra-uterine CXCL12 administration in C57BL/6 mice improved endometrial receptivity showing increased integrin β3 and its ligand osteopontin, and induced endometrial angiogenesis displaying increased numbers of vessel formation near the lining of endometrial epithelial layer with higher CD31 and CD34 expression. Furthermore, intra-uterine CXCL12 application dramatically promoted the rates of embryo implantation with no morphologically retarded embryos. Thus, our present study provides a novel evidence that improved uterine endometrial receptivity and enhanced angiogenesis induced by embryo-derived chemokine CXCL12 may aid to develop a minimally-invasive therapeutic strategy for clinical treatment or supplement for the patients with repeated implantation failure with less risk.

Morphological features of the placenta in obese women

Hypothesis/Aims of study. Obesity and severe chronic somatic pathology in a woman leads to a rapid depletion of compensatory and adaptive reserves of the placenta and to the progression of circulatory and dystrophic changes, which causes intrauterine growth retardation and reduces the likelihood of a favorable course of pregnancy and childbirth. The aim of this study was to assess the morphological features of the vascular component of placental villi in obese women. Study design, materials and methods. Histological and immunohistochemical studies were conducted on 41 placentas from obese patients with and without gestational diabetes mellitus and from healthy patients, endothelial marker CD34+ expression being assessed in chorionic villi. Results. In obese patients, chronic placental insufficiency is presented in most cases as a dissociated form with persistence of not only mature but also immature villi, which indicates early structural pathology of the placenta. Conclusion. Obesity in women contributes to more frequent chronic placental insufficiency with severe circulatory disorders and varying degrees of severity of compensatory and adaptive changes.