Mitochondrial organelle transplantation (MOT) is an innovative strategy for the treatment of mitochondrial dysfunction such as cardiac ischemic reperfusion injuries, Parkinson’s diseases, brain and spinal cord injuri...Mitochondrial organelle transplantation (MOT) is an innovative strategy for the treatment of mitochondrial dysfunction such as cardiac ischemic reperfusion injuries, Parkinson’s diseases, brain and spinal cord injuries, and amyotrophic lateral sclerosis (ALS). However, one of the major challenges for widespread usage is a methodology for preservation of isolated mitochondria. Extracellular vesicles (EVs) are phospholipid bilayer-enclosed vesicles released from cells. EVs carry a cargo of proteins, nucleic acids, lipids, metabolites, and even organelles such as mitochondria. Purpose: To test if EVs enhance the stability of isolated mitochondria. Methods: We mixed isolated mitochondria of fibroblasts with EVs of mesenchymal stromal cells (imEVs) (9:1 in volume) and stored the mixture at 2°C - 6°C for different time periods. We measured morphology, mitochondrial membrane potential (MMP) and mitochondrial ATP content at 0, 2, 5 days. Key findings: After 2 days of storage, the mito-chondria without imEVs lost approximate 70% MMP (RFU: 1822 ± 68), compared to the fresh mitochondria (RFU: 5458 ± 52) (p 0.05). In agreement with MMP, mitochondria without imEVs lost significant mitochondrial ATP content (p 0.05), after 2 days of cold storage, compared to fresh mitochondria. Microscopy showed that imEVs promoted aggregation of isolated mitochondria. Summary: The preliminary data showed that imEVs enhanced the stability of isolated mitochondria in cold storage.展开更多
Purpose: HLA-G binds to the inhibitory receptors of uterine NK cells and plays an important role in protection of fetal cells from maternal NK lysis. HLA-G also mediates tumor escape, but the immunosuppressive role is...Purpose: HLA-G binds to the inhibitory receptors of uterine NK cells and plays an important role in protection of fetal cells from maternal NK lysis. HLA-G also mediates tumor escape, but the immunosuppressive role is often neglected. These studies have focused on the examination of HLA-G expression in human breast and ovarian carcinoma and HLA-G immunosuppressive role in NK cytolysis. Methods: We examined HLA-G expression in breast and ovarian carcinoma cell lines by real time PCR, ELISA and immunofluorescent staining, and in frozen breast and ovarian carcinoma tissues by immunohistochemistry (IHC). We treated the breast cancer cell lines with anti-human HLA-G antibody or progesterone. Then, NK cytolysis was measured by using MTT assay. Results: We find breast and ovarian cancer cell lines increase the expression of HLA-G mRNA and protein, compared to normal cells. IHC shows that 100% of frozen breast and ovarian carcinoma tissues overexpress HLA-G protein. HLA-G IHC scores of breast and ovarian carcinoma are significantly higher than normal breast and ovarian tissues, respectively (both p < 0.01). Blocking HLA-G of the breast cancer cells by the antibody increases NK cytolysis. Progesterone upregulates HLA-G mRNA and protein of human breast cancer cell lines. The increased HLA-G expression by progesterone suppresses the NK cytolysis. Conclusion: Human breast and ovarian carcinoma overexpress HLA-G immunosuppressive molecules. Blocking HLA-G protein by antibody improves the cytolysis of NK cells against human breast cancer cell lines. In contrast, upregulation of HLA-G expression by progesterone impairs NK cytolytic function. Thus, HLA-G is a new immune checkpoint protein and potential cancer immunotherapeutic target.展开更多
Objective: The purpose of initiating contrast enhanced digital mammography in our center was to evaluate the complimentary benefit of this technology with screening digital mammography and real time ultrasound in equi...Objective: The purpose of initiating contrast enhanced digital mammography in our center was to evaluate the complimentary benefit of this technology with screening digital mammography and real time ultrasound in equivocal cases and high risk patients with dense breast. The intended goal was to reduce the incidence of further diagnostic and invasive procedures. Methods: Patients thought to be candidates who had good renal function confirmed by serum Blood Urea Nitrogen (BUN) and Creatinine were offered the procedure, and 225 patients had the procedure during the period of March 2013 through November 2014. The contrast enhanced digital mammograms (SenoBright) are performed on the Senograph Essential Unit. A total of 8 images are obtained: 4 conventional digital mammograms and 4 contrast enhanced digital mammograms. The patients with a positive SenoBright study had a tissue diagnosis of the lesion obtained by either a stereotactic needle biopsy, ultrasound guided core biopsy, or ultrasound directed open excisional biopsy. Results: The 225 patients who had the procedure included high risk patients with dense breast (41), patients with abnormal mammograms (92), and patients with equivocal clinical, mammographic and real time ultrasound findings (92). 31 studies were interpreted as positive and 194 as negative. 33 biopsies were performed, with 31 patients having a positive study and 2 patients with a negative study. 22 cancers were detected. Conclusion: We found that the addition of dynamic contrast enhanced digital subtraction mammography (SenoBright) was helpful in distinguishing malignant from non-malignant lesions. It was also effective in demonstrating multifocal lesions and identifying non-palpable occult carcinomas in the dense breast. It proved to be a valuable complimentary adjunctive diagnostic modality for a comprehensive clinical breast center.展开更多
文摘Mitochondrial organelle transplantation (MOT) is an innovative strategy for the treatment of mitochondrial dysfunction such as cardiac ischemic reperfusion injuries, Parkinson’s diseases, brain and spinal cord injuries, and amyotrophic lateral sclerosis (ALS). However, one of the major challenges for widespread usage is a methodology for preservation of isolated mitochondria. Extracellular vesicles (EVs) are phospholipid bilayer-enclosed vesicles released from cells. EVs carry a cargo of proteins, nucleic acids, lipids, metabolites, and even organelles such as mitochondria. Purpose: To test if EVs enhance the stability of isolated mitochondria. Methods: We mixed isolated mitochondria of fibroblasts with EVs of mesenchymal stromal cells (imEVs) (9:1 in volume) and stored the mixture at 2°C - 6°C for different time periods. We measured morphology, mitochondrial membrane potential (MMP) and mitochondrial ATP content at 0, 2, 5 days. Key findings: After 2 days of storage, the mito-chondria without imEVs lost approximate 70% MMP (RFU: 1822 ± 68), compared to the fresh mitochondria (RFU: 5458 ± 52) (p 0.05). In agreement with MMP, mitochondria without imEVs lost significant mitochondrial ATP content (p 0.05), after 2 days of cold storage, compared to fresh mitochondria. Microscopy showed that imEVs promoted aggregation of isolated mitochondria. Summary: The preliminary data showed that imEVs enhanced the stability of isolated mitochondria in cold storage.
文摘Purpose: HLA-G binds to the inhibitory receptors of uterine NK cells and plays an important role in protection of fetal cells from maternal NK lysis. HLA-G also mediates tumor escape, but the immunosuppressive role is often neglected. These studies have focused on the examination of HLA-G expression in human breast and ovarian carcinoma and HLA-G immunosuppressive role in NK cytolysis. Methods: We examined HLA-G expression in breast and ovarian carcinoma cell lines by real time PCR, ELISA and immunofluorescent staining, and in frozen breast and ovarian carcinoma tissues by immunohistochemistry (IHC). We treated the breast cancer cell lines with anti-human HLA-G antibody or progesterone. Then, NK cytolysis was measured by using MTT assay. Results: We find breast and ovarian cancer cell lines increase the expression of HLA-G mRNA and protein, compared to normal cells. IHC shows that 100% of frozen breast and ovarian carcinoma tissues overexpress HLA-G protein. HLA-G IHC scores of breast and ovarian carcinoma are significantly higher than normal breast and ovarian tissues, respectively (both p < 0.01). Blocking HLA-G of the breast cancer cells by the antibody increases NK cytolysis. Progesterone upregulates HLA-G mRNA and protein of human breast cancer cell lines. The increased HLA-G expression by progesterone suppresses the NK cytolysis. Conclusion: Human breast and ovarian carcinoma overexpress HLA-G immunosuppressive molecules. Blocking HLA-G protein by antibody improves the cytolysis of NK cells against human breast cancer cell lines. In contrast, upregulation of HLA-G expression by progesterone impairs NK cytolytic function. Thus, HLA-G is a new immune checkpoint protein and potential cancer immunotherapeutic target.
文摘Objective: The purpose of initiating contrast enhanced digital mammography in our center was to evaluate the complimentary benefit of this technology with screening digital mammography and real time ultrasound in equivocal cases and high risk patients with dense breast. The intended goal was to reduce the incidence of further diagnostic and invasive procedures. Methods: Patients thought to be candidates who had good renal function confirmed by serum Blood Urea Nitrogen (BUN) and Creatinine were offered the procedure, and 225 patients had the procedure during the period of March 2013 through November 2014. The contrast enhanced digital mammograms (SenoBright) are performed on the Senograph Essential Unit. A total of 8 images are obtained: 4 conventional digital mammograms and 4 contrast enhanced digital mammograms. The patients with a positive SenoBright study had a tissue diagnosis of the lesion obtained by either a stereotactic needle biopsy, ultrasound guided core biopsy, or ultrasound directed open excisional biopsy. Results: The 225 patients who had the procedure included high risk patients with dense breast (41), patients with abnormal mammograms (92), and patients with equivocal clinical, mammographic and real time ultrasound findings (92). 31 studies were interpreted as positive and 194 as negative. 33 biopsies were performed, with 31 patients having a positive study and 2 patients with a negative study. 22 cancers were detected. Conclusion: We found that the addition of dynamic contrast enhanced digital subtraction mammography (SenoBright) was helpful in distinguishing malignant from non-malignant lesions. It was also effective in demonstrating multifocal lesions and identifying non-palpable occult carcinomas in the dense breast. It proved to be a valuable complimentary adjunctive diagnostic modality for a comprehensive clinical breast center.
