Latest research and prospect of CD47 in kidney transplantation / 器官移植

CD47 is a transmembrane protein widely expressed on cell surface, which is considered as a key molecule for immune escape. With an increasing number of related studies, the role of CD47 and its ligands in immunomodulatory effects has been gradually understood. Recent studies have investigated the role of CD47 in ischemia-reperfusion injury of allogenetic kidney transplantation, rejection and xenotransplantation. Nevertheless, the specific role and the key mechanism remain elusive. In this article, the structure and function of CD47, common CD47 ligands, the relationship between CD47 and kidney transplantation, and the application of CD47 in kidney transplantation were reviewed, the latest research progress of CD47 in kidney transplantation was summarized, and the limitations of current research and subsequent research direction were analyzed, aiming to provide reference for subsequent application of CD47 in allogeneic and kidney xenotransplantation.

The prognostic effect of immunohistochemical staining rates in patients with non–muscle-invasive bladder cancer

Context: Despite the follow-up protocols developed in non–muscle-invasive bladder cancer patients, progression and recurrence could not be prevented. Aims: We aimed to investigate whether proteins such as OCT-4, CD47, p53, Ki-67, and Survivin, which increase in bladder cancer cells, can be used as prognostic markers for patients with non–muscle-invasive bladder cancer. Settings and Design: The study included a total of 89 patients with newly diagnosed non–muscle-invasive bladder cancer between January 2015 and December 2020. Materials and Methods: Levels of OCT-4, CD47, p53, K?-67, and Survivin proteins in cancer cells were determined with a semi-quantitative immunohistochemical experiment. Pathological data and survival rates were compared according to the staining rates. Statistical Analysis Used: Data obtained in the study were analyzed statistically with SPSS 22.0 (SPSS, Chicago, IL, USA). Results: The mean age of the patients was 64.25 ± 9.91 years, and the median follow-up period was 55 months. Recurrence rate was determined to be 36% (n = 32), and the rate of progression at 40.4% (n = 36). The staining rates were stronger for each marker in the progression group and advanced-stage tumors (p < 0.001). The findings of the multivariate analysis carried out as part of the study showed that older age and higher tumor stage were independent risk factors for recurrence-free survival (HR = 1.048 and 7.074, respectively; P = 0.02). Also, higher tumor stages, diameters, and grades were associated with reduced progression-free survival (HR = 0.105, 0.395, 0.225, respectively; P < 0.05). Conclusions: Although immunohistochemical staining rates are promising, it is more appropriate to use tumor characteristics when assessing survival rate in patients with non–muscle-invasive bladder cancer.

Radix Tetrastigme Polysaccharide Promotes Antitumor Immune Response in Lewis Lung Cancer Mice / 中国肺癌杂志

BACKGROUND@#Lung cancer has a high incidence and mortality rate, but the treatment of lung cancer still lacks low toxicity and efficient anti-tumor drugs. Polysaccharide from radix tetrastigme has development value in anti-tumor treatment methods. This study was to observe the effect of polysaccharide from radix tetrastigme on immune response of Lewis lung cancer mice and explore its molecular mechanism.@*METHODS@#Lewis lung cancer mouse models were established and randomly grouped. The spleen polypeptide group was intragastric with 50 mg/kg spleen polypeptide, and the radix tetrastigme polysaccharide low, medium and high dose groups were intragastric with 62.5, 125 and 250 mg/kg radix tetrastigme polysaccharide, respectively, and the model group and the control group were intragastric with equivolume normal saline. Tumor formation and metastasis were compared. Haematoxylin-eosin (HE) staining was used to observe the pathological changes of tumor cells. Macrophage phagocytosis, apoptosis, M1/M2 polarization, T cell subsets and cytokine levels in peripheral blood were detected by flow cytometry. The proliferation activity of macrophages was detected by methyl thiazolyldiphenyl tetrazolium (MTT) assay. Dendritic cell (DC) antigen presenting function was detected by chlorophenol red-β-D-galactopyranoside (CPRG) method. Tumor tissue differentiation antigen cluster 47 (CD47) mRNA and protein expression and macrophage signal regulatory protein α (SIRRP α) expression were detected by real time quantitative polymerase chain reaction (RT-qPCR) and Western blot (WB).@*RESULTS@#The tumor inhibition rates and anti-metastasis rates in the 3-dose radix tetrastigme polysaccharide group and the spleen polypeptide group were higher than those in the model group, and the pathological injury of tumor tissue were severer, and the positive rate of phagocytosis of ink by macrophages and the efficiency of phagocytosis of tumor cells were increased; the apoptosis rate of macrophages was decreased; the proliferation activity of macrophages, polarization ratio of macrophages to M1 type, DC antigen presenting ability, CD4+, CD4+/CD8+ levels were increased; the level of serum tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and the expression of tumor tissue CD47, macrophage SH2-containing protein tyrosine phosphatase 1 (SHP-1), SH2-containing protein tyrosine phosphatase 2 (SHP-2), and phosphorylation signal regulatory protein α (p-SIRPα) were decreased, and the differences were statistically significant (P<0.05). There were no significant differences in the above indexes between low-dose radix tetrastigme polysaccharide group and spleen polypeptide group (P>0.05), and the effects of radix tetrastigme polysaccharide were dose-dependent.@*CONCLUSIONS@#Radix tetrastigme polysaccharide can inhibit tumor growth, metastasis and immune response in Lewis lung cancer mice, and its mechanism may be related to inhibiting SIRP/CD47 signaling pathway.

Mitigating of the interference of anti-CD47 monoclonal antibody on transfusion compatibility detection / 中国输血杂志

【Objective】 To evaluate the interference of anti-CD47 monoclonal antibody on transfusion compatibility detection, in order to establish methods for removing interference and evaluate its efficacy. 【Methods】 Blood samples from 8 patients in our clinical trial who were treated with anti-CD47 monoclonal antibody from Tianjing and Xinda were collected. ABO and Rh blood group antigen identification, direct anti-human globulin test, unexpected antibody screening test and cross-matching test were performed by ZZAP, Gamma-clone(an anti-globulin reagent lacking IgG4) and Immucor Capture-R solid phase agglutination kit. 【Results】 ABO blood group identification of 5 subjects were interfered after treatment with anti-CD47 monoclonal antibody. All 8 subjects showed 2+ to 4+ agglutination intensity on direct anti-human globulin test and 3+ to 4+ on unexpected antibody screening. The results of unexpected antibody screening by Gamma-clone and Immucor Capture-R solid phase agglutination kit were all negative, while the cross-matching test were compatible. Patients with anemia caused by CD47 monoclonal antibody treatment were transfused with 2 U suspension red blood cells, and the evaluation showed that the transfusion was effective. 【Conclusion】 The CD47 monoclonal antibody can interfere with transfusion compatibility detection, and the use of antiglobulin reagents lacking IgG4 and Immucor Capture-R solid phase agglutination kit can remove the interference, with good transfusion efficacy in patients.

CD47 blockade improves the therapeutic effect of osimertinib in non-small cell lung cancer / 医学前沿

The third-generation epidermal growth factor receptor (EGFR) inhibitor osimertinib (OSI) has been approved as the first-line treatment for EGFR-mutant non-small cell lung cancer (NSCLC). This study aims to explore a rational combination strategy for enhancing the OSI efficacy. In this study, OSI induced higher CD47 expression, an important anti-phagocytic immune checkpoint, via the NF-κB pathway in EGFR-mutant NSCLC HCC827 and NCI-H1975 cells. The combination treatment of OSI and the anti-CD47 antibody exhibited dramatically increasing phagocytosis in HCC827 and NCI-H1975 cells, which highly relied on the antibody-dependent cellular phagocytosis effect. Consistently, the enhanced phagocytosis index from combination treatment was reversed in CD47 knockout HCC827 cells. Meanwhile, combining the anti-CD47 antibody significantly augmented the anticancer effect of OSI in HCC827 xenograft mice model. Notably, OSI induced the surface exposure of "eat me" signal calreticulin and reduced the expression of immune-inhibitory receptor PD-L1 in cancer cells, which might contribute to the increased phagocytosis on cancer cells pretreated with OSI. In summary, these findings suggest the multidimensional regulation by OSI and encourage the further exploration of combining anti-CD47 antibody with OSI as a new strategy to enhance the anticancer efficacy in EGFR-mutant NSCLC with CD47 activation induced by OSI.

Building on the backbone of CD47-based therapy in cancer: Combination strategies, mechanisms, and future perspectives

Described as a "don't eat me" signal, CD47 becomes a vital immune checkpoint in cancer. Its interaction with signal regulatory protein alpha (SIRPα) prevents macrophage phagocytosis. In recent years, a growing body of evidences have unveiled that CD47-based combination therapy exhibits a superior anti-cancer effect. Latest clinical trials about CD47 have adopted the regimen of collaborating with other therapies or developing CD47-directed bispecific antibodies, indicating the combination strategy as a general trend of the future. In this review, clinical and preclinical cases about the current combination strategies targeting CD47 are collected, their underlying mechanisms of action are discussed, and ideas from future perspectives are shared.

miR-29c-3p represses the angiogenesis of esophageal squamous cell carcinoma by targeting SERPINH1 to regulate the Wnt signaling pathway

Purpose: Esophageal squamous cell carcinoma (ESCC) is characterized by early metastasis and late diagnosis. miR-29c-3p is confirmed to repress angiogenesis in multiple tumor types. Yet, the functions of miR-29c-3p in the mechanism of ESCC angiogenesis, which were not sufficiently explored previously, were exactly what we investigated here at the molecular level. Methods: The mRNA level of miR-29c-3p and Serpin peptidase inhibitor clade H member 1 (SERPINH1) in ESCC tissues were assessed via bioinformatics analysis. Thereafter, miR-29c-3p and SERPINH1 (HSP47) mRNA level in ESCC cell lines was evaluated via quantitative real-time polymerase chain reaction. The effects of abnormal miR-29c-3p and SERPINH1 expression on ESCC cell viability, proliferation, migration, invasion, and HUVEC angiogenesis were examined via CCK8, colony formation, transwell, and angiogenesis assays, respectively. The protein levels of SERPINH1, vascular endothelial growth factor-A (VEGFA), Wnt-1, ?-catenin, and p-?-catenin were evaluated via Western blot. Expression of VEGFA secreted by ESCC cells was measured via enzyme-linked immunosorbent assay. Treatment with the Wnt activator BML-284 further revealed the way miR-29c-3p mediated the Wnt signaling pathway and its effects on angiogenesis. Results: Herein, we revealed a decrease of miR-29c-3p expression in ESCC tissues and cells, while the overexpressed miR-29c-3p could remarkably suppress ESCC cell progression, as well as HUVEC angiogenesis. Meanwhile, overexpressed miR-29c-3p notably downregulated VEGFA and repressed the Wnt signaling pathway. Treatment with the Wnt activator BML-284 could reverse the inhibition of HUVEC angiogenesis caused by miR-29c-3p. SERPINH1 was a downstream target of miR-29c-3p. SERPINH1 knockdown suppressed the malignant phenotypes of ESCC cells and impeded the Wnt signaling activation, while such suppression was reversed through miR-29c-3p inhibitor. Conclusions: We confirmed the mechanism that miR-29c-3p targeted SERPINH1, thus regulating angiogenesis in ESCC through the Wnt signaling pathway. It improves the understanding of angiogenesis in ESCC and offers new ideas for the research of ESCC treatment strategies in the future.

Research Advances on CD47 Molecules in Tumor Microenvironment of Diffuse Large B-cell Lymphoma / 肿瘤防治研究

Diffuse large B-cell lymphoma (DLBCL) is a common, highly aggressive and heterogeneous hematologic malignancy in adults. Patients with DLBCL have substantially differences in molecular biological characteristics, clinical manifestations, and prognosis. Increasing evidence shows that the tumor microenvironment plays an important role in the occurrence and development of DLBCL. CD47, an integrin related protein, is overexpressed in DLBCL cells and plays a key role in immune escape of lymphoma. This work reviews the research progress of CD47 in DLBCL TME in terms of CD47-related signal pathway, CD47 role in DLBCL TME, and therapeutic strategies targeting CD47 in DLBCL TME.

Significance of CD47 expression in endometrial carcinoma

Objectives: CD47 is a membrane protein that belongs to the immunoglobulin superfamily and regulates macrophage phagocytosis negatively. As CD47 expression at the cancer cell membrane would inhibit the phagocytic activity of immune cells, it is connected to an unfavorable prognosis in leukemia and malignancies of various solid organs. Materials and Methods: In this study, retrospectively evaluated 72 patients who had been diagnosed with endometrial carcinoma at Pathology Department and had undergone total abdominal hysterectomy and bilateral salpingo-oophorectomy (TAH + BSO) and/or lymphadenectomy. CD47 expression was evaluated in tumorous and nontumor areas in all patients considering cytoplasmic and membranous brown staining in cells. The proportion of expression was evaluated as well as the intensity and an “h score” was obtained. This score was compared with known prognostic parameters. Results: CD47 expressions showed a statistically significant correlation with tumor grade (P < 0.05); however, no significant relationship was observed with myometrial invasion depth and lymph vascular invasion status (P = 0.923 and P = 0.754, respectively). Conclusions: As with other tumors, anti-CD47 antibody may be an alternative treatment option in patients with high-grade endometrial carcinoma.

Progress of heat shock protein 47 in glioma / 肿瘤研究与临床

Heat shock protein 47 (HSP47) is mainly involved in regulating collagen folding, secretion and maturation in the tumor microenvironment (TME) and is widely amplified in human cancers. HSP47 has been shown to be overexpressed in a variety of extracranial tumors. In glioma, the expression of HSP47 correlates with the grading of glioma and is involved in proliferation, invasion, angiogenesis, and immune regulation of glioma, regulates the TME of glioma, and promotes the survival of glioma stem cells, which may be related to the heterogeneity of glioma. This article reviews the progress of HSP47 in the development and progression of glioma, and discusses the significances of HSP47 in the proliferation, invasion, targeted therapy and immunotherapy of glioma.

The protective effect of inhibiting CD47 expression on radiation-induced lung injury / 中华放射肿瘤学杂志

Objective:To study the protective anti-radiation effect of inhibiting CD47 expression in the lung tissues of mice and to explore the associated mechanism.Methods:Female C57BL/6 mice ( n = 60) were randomly divided into four groups: normal, blank, negative and positive. The blank group received only whole-lung irradiation; The negative group received whole-lung irradiation and tracheal infusion of adeno-associated virus containing nonsense sequence shRNA; The positive group received whole lung irradiation and tracheal drip containing adeno-associated virus containing shRNA expression of CD47. Fresh blood samples were collected at 24 h, 4 weeks, and 12 weeks post-irradiation, respectively. The expression level of CD47 mRNA was determined by RT-PCR. Determination of hydroxyproline content by alkaline hydrolysis. The LC3 expression level was measured by immunohistochemical staining. Serum transforming growth factor-β 1 (TGF-β 1) and tumor necrosis factor-α (TNF-α) levels were assessed by ELISA. Results:RT-PCR showed that the relative expression level of CD47 mRNA in the lung tissues in the positive group was significantly lower compared to those in the negative group, the normal group, the blank group (24-hour, P were <0.001,<0.001,<0.001, respectively. 4-weeks, P were <0.001,0.003,0.001, respectively. 12-weeks, P were 0.009, 0.002, 0.005, respectively). There were no significant differences in CD47 mRNA expression in the three groups except the positive group (all P>0.05), and there was no significant difference in CD47 mRNA expression with time in each group (all P>0.05).The serum TGF-β1 content was higher in the 24 h, 4-week, and 12-week blank groups ( P were <0.001, 0.003 and 0.003, respectively) and negative groups( P were 0.001, 0.021 and 0.034, respectively) after irradiation than that in the mice in the normal group. At the same time, the serum TNF-α of positive group after irradiation (24 hours, 4 weeks, 12 weeks, P were 0.022, <0.001, <0.001, respectively) were significantly higher than those of the normal group. The content of hydroxyproline in the blank group was significantly higher than that in the normal group (4 weeks, 12 weeks, P were 0.002, <0.001, respectively). Immunohistochemical indications: 24 h after irradiation was higher than the expression of LC3 in mouse lung tissue at 4 weeks and 12 weeks (all P < 0.001). The difference between the negative group and the blank group was not obvious ( P>0.05). Conclusions:Inhibition of CD47 expression can reduce the degree of radiation-induced pneumonia and pulmonary fibrosis probably via enhanced autophagy. CD47 may represent a novel target for the protection of radiation-induced lung injury.

Application of CD47 in targeted therapy strategies for hematologic neoplasms / 白血病·淋巴瘤

CD47 is widely expressed on the cell surface, and combines with signal regulatory protein α (SIRPα) to transmit the "don't eat me" signal, which plays a key role in self-recognition and tumor immune escape. Studies have shown that the high expression of CD47 in different hematologic neoplasms is associated with the occurrence, progression and poor prognosis of tumors. As a new immune checkpoint, CD47 is gradually becoming an effective target for tumor immunotherapy. and various related preclinical and clinical studies for hematologic neoplasms are underway. This article summarizes the application of CD47 in hematologic neoplasms, in order to provide references for the treatment.

Study on the mechanism of curcumol inhibiting the proliferation of breast cancer cells T 47D / 中国药房

OBJECTIVE To study the mechanism of curcumol inhibiting the pro liferation of breast cancer cells T 47D. METHODS MTT assay was used to detect the inhibitory effects of different doses of curcumol （0，6.25，12.5，25，50，100 μg/mL）on the proliferation of T 47D cells. After treated with curcumol （12.5，25，50，100 μg/mL），the morphology of T 47D cells was observed by inverted phase contrast microscope. The cell cycle and the levels of reactive oxygen species （ROS）were detected by flow cytometry. Quantitative real-time PCR （qRT-PCR）was used to detect the expressions of proliferating cell nuclear antigen （PCNA），cell cycle regular p 21 and cyclin-dependent kinase 2（CDK2）mRNA. Western blot assay was used to detect the protein expression of CDK 2，CDK6，Cyclin D ，PCNA，nucler transcription factor E 2-related factor （Nrf2）and Kelch-like ECH associated protein 1（Keap1）. Breast cancer cells T 47D were divided into 2 groups，one group was given different doses of curcumol ，and another group was given curcumol 33 μg/mL for 6，12，24，48 h. After the optimal oxidation time and administration concentration were determined according to the results of the above two groups ，the blank control group ，N-acetylcysteine（NAC）group（ROS antioxidant NAC alone ），curcumol group （curcumol alone ），curcumol combined with NAC group （pretreatment with ROS antioxidant NAC ，and then adding into curcumol ）. Cell cycle and fluorescence intensity of ROS were detected. RESULTS Curcumol could significantly increase the inhibitory rate of the proliferation of T 47D cells （P＜0.05 or P＜0.01），and showed a certain dose and time dependent trend. Curcumol blocked the ， cycle in the G 1 phase and significantly increased the level of ROS （P＜0.05 or P＜0.01）；ROS antioxidant NAC could significantly reverse above inductive effect of curcumol （P＜ 0.01）. qRT-PCR showed that curcumol down-regulated the com expression of PCNA and CDK 2 mRNA and up-regulated the expression of p 21 mRNA（P＜0.05 or P＜0.01）. Western blot assay showed that curcumol significantly down-regulated the edu.cn protein expression of Keap 1，Nrf2，CDK2，CDK6 and Cyclin D（P＜0.05，P＜0.01）；ROS antioxidant NAC could reverse the down-regulation effects of curcumol on the expression of these proteins（P＜0.05 or P＜0.01）. CONCLUSIONS Curcumol may induce oxidative stress and cell arrest in G 1 phase to inhibit the proliferation of T 47D cells.

Interfering with B7-H4 expression can inhibit proliferation of breast cancer cells by down-regulating E2F family related transcription factors / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨干扰B7-H4表达对乳腺癌细胞增殖、凋亡、周期以及相关下游分子表达的影响。方法：利用脂质体转染技术分别将特异性靶向B7-H4的siRNA（siB7-H4）及其阴性对照（siNC）转染至对数生长期的乳腺癌T47D和MCF-7细胞，分别命名为T47D-siB7-H4、T47D-siNC、MCF-7-siB7-H4和MCF-7-siNC组。用qPCR法和WB法验证siRNA干扰效果及其对细胞周期分子cyclin D1表达的影响，CCK-8法和FCM分别检测干扰B7-H4表达对T47D和MCF-7细胞增殖、周期和凋亡的影响，qPCR法检测B7-H4干扰对E2F家族相关转录因子表达的影响。结果：成功构建干扰B7-H4表达的乳腺癌T47D和MCF-7细胞。与T47D-siNC和MCF-7-siNC组相比，T47D-siB7-H4和MCF-7-siB7-H4组细胞中B7-H4 mRNA和蛋白表达水平均显著降低、细胞增殖能力显著降低（均P<0.01），并伴有G1/S期细胞周期阻滞以及cyclin D1表达下调（均P<0.01），但细胞凋亡率差异无统计学意义（均P>0.05）。与T47D-siNC相比，干扰B7-H4后T47D细胞中E2F1、E2F2、E2F7和E2F8 mRNA水平有不同程度的降低（均P<0.01）；与MCF-7-siNC相比，干扰B7-H4后MCF-7细胞中E2F1、E2F2、E2F3、E2F7和E2F8 mRNA水平均有不同程度的降低（P<0.05或P<0.01）。结论：干扰乳腺癌细胞B7-H4表达可下调cyclin D1和E2F家族相关转录因子的表达，导致细胞周期阻滞并抑制细胞增殖。

TRIM47 promotes ovarian cancer cell proliferation, migration, and invasion by activating STAT3 signaling

Abstract Objectives Tripartite Motif 47 (TRIM47) protein plays a prominent role in many cancers. This study aimed to investigate the biological roles of TRIM47 in ovarian cancer. Methods TRIM47 was knocked down and overexpressed in ovarian cancer cell lines SKOV3 and OVCAR3, and the effects on proliferation, clone formation, apoptosis, invasion, and growth of xenograft tumors in nude mice were determined. The expression levels of the selected candidates were tested by western blotting and quantitative real-time PCR. Results TRIM47 knockdown suppressed proliferation and encourages apoptosis of ovarian cancer cells. Similarly, TRIM47 knockdown suppressed ovarian cancer cell invasion, migration, and epithelial-mesenchymal transition. Ovarian cancer cell xenograft assays demonstrated that TRIM47 knockdown significantly inhibited tumor growth. Mechanistically, TRIM47 knockdown suppressed STAT3 phosphorylation and the expression of several downstream genes, including MCL-1, MMP2, and c-MYC. Silencing of STAT3 partially prevented TRIM47-induced tumor cell proliferation and invasion. Conclusion The present study's findings demonstrate that by activating STAT3 signaling, TRIM47 functions as an oncogene in ovarian cancer. TRIM47, therefore, appears to be a potential target for ovarian cancer prevention and/or therapy.

Research Progress on Relation Between CD47 and Immunotherapy on Lymphoma / 肿瘤防治研究

CD47 is a member of the immunoglobulin superfamily. It is expressed on various cells and tissues of human, but it is more expressed on tumor cells, especially in various hematopoietic tumors. The combination of CD47 expressed on tumor cells with signal regulator protein α (SIRPα) on macrophages inhibits the phagocytosis of tumors by macrophages. The reaction can lead to tumor immune escape. CD47 has become a new hot spot in tumor research. This article reviews the correlation between the structure and expression of CD47, CD47-SIRPα, CD47-targeting antibody drugs and lymphoma immunotherapy.

Development and validation of ELISA method for detection of human signal regulatory protein α-anti-CD20 mouse chimeric antibody fusion protein IMM0306 in human serum / 药学学报

IMM0306 is a recombinant human signal regulatory protein α-anti-CD20 mouse chimeric antibody fusion protein, intended to treat refractory or recurrent CD20 positive B-cell non-Hodgkin lymphoma (B-NHL) in clinical. In this study, an ELISA method was established to evaluate the pharmacokinetics of IMM0306 in human serum. The experiment was approved by the Ethics Committee of the Cancer Hospital of the Chinese Academy of Medical Sciences (No. CTR20192612). Recombinant human CD47 protein was coated with the plate overnight, blocking the plate with 5% skin milk for 2 h. After washing, 100 μL per well standard and unknown samples were added, and incubated for 1.5 h. After washing, the detection antibody Anti-IMM0306-Biotin was added and incubated for 1 h, and then HRP-labeled streptavidin was added for 1 h, and the color was detected. The optimal concentration of coating reagent was 2 μg·mL-1 by ELISA method, and the optimal dilution of anti-IMM0306-biotin and SA-HRP were 1∶500 and 1∶5 000, respectively. The lower limit of quantitation was 4 ng·mL-1, and the standard curve range was 4-100 ng·mL-1. The verification results of the method meets the corresponding acceptance criteria, and can be used in IMM0306 clinical pharmacokinetic studies.

miR-1207-5p regulates the proliferation, migration and invasion of breast cancer T47D stem cells by targeting LIMD1 / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨miR-1207-5p对乳腺癌T47D干细胞增殖、迁移和侵袭的影响及其可能的机制。方法: 以IGF-1、EGF、bFGF诱导、富集乳腺癌T47D干细胞并进行成球培养，流式细胞术分离干细胞，采用WB法检测干细胞分子标志物。采用qPCR检测干细胞中miR-1207-5p的表达水平，双荧光素酶报告基因实验分析miR-1207-5p和LIMD1的靶向关系。CCK-8、Transwell和划痕实验检测T47D干细胞的增殖、迁移和侵袭能力。WB法检测干细胞中LIMD1蛋白的表达水平。结果: 分离的T47D干细胞能够形成细胞球，细胞球体积随培养天数的增加而增加；干细胞分子标志物ALDH1、ESA和OCT4的表达水平较亲本T47D细胞显著升高（P<0.05或P<0.01），miR-1207-5p在干细胞中高表达（P<0.01）。过表达miR-1207-5p显著促进T47D干细胞的增殖、迁移和侵袭（均P<0.01），敲降miR-1207-5p显著抑制T47D干细胞的增殖、迁移和侵袭（均P<0.01）。miR-1207-5p靶向下调LIMD1的表达（P<0.01），miR-1207-5p通过靶向下调LIMD1促进乳腺癌T47D干细胞的增殖、迁移和侵袭能力（P<0.05或P<0.01）。结论： miR-1207-5p通过靶向下调LIMD1的表达来促进乳腺癌T47D干细胞的增殖、迁移和侵袭能力。

Ultra-short-course and intermittent TB47-containing oral regimens produce stable cure against Buruli ulcer in a murine model and prevent the emergence of resistance for

Buruli ulcer (BU), caused by

Monoclonal anti-CD47 interference in pre-transfusion test and mitigation measures, a case report / 中国输血杂志

【Objective】 To discuss the interference of anti-CD47 in pre-transfusion test and the mitigation measures. 【Methods】 Blood sample of one patient received anti-CD47 treatment was collected to conduct routine serological tests including ABO/Rh phenotype, direct anti-human globulin test, irregular antibody screening, antibody identification and cross-match. Packed platelet from multiple type O blood donors was used to absorb with patient′s plasma. The patient′s plasma was absorbed with CCDee, ccDEE and ccdee red cells, respectively. Anti-IgG monoclonal Gamma-clone which lacks reactivity with human subclass IgG4 was used to perform antibody screening and cross-match. Capture-R was used to perform antibody screening. 【Results】 The direct anti-human globulin test was positive(1+ ), the reactivity in all phases was strong positive(3+ -4+ ). The anti-CD47 was eliminated after platelet and red cells absorption. Antibody screening became negative using Gamma-clone and Capture-R, and cross-match successfully using Gamma-clone. 【Conclusion】 Anti-CD47 monoclonal antibody can interfere with pre-transfusion test and cross matching. To remove the interference of anti-CD47 requires the use of Gamma-clone anti-IgG in the indirect antiglobulin testing or Capture-R.