Mechanism and intervention of murine transfusion-related acute lung injury caused by anti-CD36 antibodies.

Anti-CD36 Abs have been suggested to induce transfusion-related acute lung injury (TRALI) upon blood transfusion, particularly in Asian populations. However, little is known about the pathological mechanism of anti-CD36 Ab-mediated TRALI, and potential therapies have not yet been identified. Here, we developed a murine model of anti-CD36 Ab-mediated TRALI to address these questions. Administration of mouse mAb against CD36 (mAb GZ1) or human anti-CD36 IgG, but not GZ1 F(ab')2 fragments, induced severe TRALI in Cd36+/+ male mice. Predepletion of recipient monocytes or complement, but not neutrophils or platelets, prevented the development of murine TRALI. Moreover, plasma C5a levels after TRALI induction by anti-CD36 Abs increased more than 3-fold, implying a critical role of complement C5 activation in the mechanism of Fc-dependent anti-CD36-mediated TRALI. Administration of GZ1 F(ab')2, antioxidant (N-acetyl cysteine, NAC), or C5 blocker (mAb BB5.1) before TRALI induction completely protected mice from anti-CD36-mediated TRALI. Although no significant amelioration in TRALI was observed when mice were injected with GZ1 F(ab')2 after TRALI induction, significant improvement was achieved when mice were treated postinduction with NAC or anti-C5. Importantly, anti-C5 treatment completely rescued mice from TRALI, suggesting the potential role of existing anti-C5 drugs in the treatment of patients with TRALI caused by anti-CD36.

Effects of message framing on recruiting Rh-D-negative blood donors in an emergency situation: Two randomized trials.

OBJECTIVES: Prospect theory categorizes message framing according to whether it emphasizes a possible loss or a possible gain. Promotions of blood donation worldwide mainly focus on gain-framed appeal. The objective of the current study is to examine the effects of message framing on recruiting Rh-D-negative blood donors (RDNBDs) in an emergency situation. DESIGN: Two randomized trials. METHODS: In Study 1, 813 group O and 500 group B RDNBDs were randomly assigned to three groups receiving three different recruitment text messages: (1) gain-framed, (2) loss-framed, and (3) information messages. In addition, 613 group A and 148 group AB RDNBDs were marked as (4) no message group. In Study 2, 758 RDNBDs were randomly provided one of two versions of materials focusing on either the possible survival (gain-framed) or the death (loss-framed) of a Rh-D-negative patient needed a blood transfusion. These participants then completed a questionnaire to examine the possible mechanisms underlying the observed effects. RESULTS: Compared to not receiving any message, significantly more RDNBDs re-donated within 14 days after they received a loss-framed message. Study 2 found that RDNBDs who read the loss-framed material expressed more willingness to donate immediately than those who read the gain-framed material. RDNBDs with high-risk perception expressed a greater blood donation intention in the loss-framed group. The loss-framed message made participants perceive others' needs more strongly, which generated more empathy, and thus increased blood donation intention. CONCLUSIONS: The results suggested that recruiting RDNBDs using a loss-framed message is suitable under an emergency.

Blood donor recruitment in Guangzhou, China, during the 2019 novel coronavirus (COVID-19) epidemic.

BACKGROUND: The coronavirus disease 2019 (COVID-19) epidemic affected blood collection in Guangzhou, China. STUDY DESIGN AND METHODS: This paper includes three studies. The observational study reported the trends of blood collection during the epidemic in Guangzhou, China. The cross-sectional survey investigated factors influencing blood donation during the COVID-19 epidemic, and a self-administered questionnaire was given to 1584 street whole blood donors (SWBDs) who donated during the epidemic. The randomized controlled trial involved 19 491 SWBDs who donated in 2019 but did not donate during the epidemic. Trial participants were randomly assigned to two intervention groups: Group 1 completed Questionnaire 1, which contained precautionary measures in response to COVID-19 and other messages about blood donation during the epidemic; Group 2 completed Questionnaire 2, which did not include this information. A control group did not receive any questionnaire. RESULTS: As measures were implemented, the number of blood donors increased accordingly. Both first-time and repeat SWBDs perceived the same level of blood need and donated blood because it would save lives. SWBDs who completed Questionnaire 1 expressed a greater intention to donate during the epidemic. Enabling blood donors to perceive a higher level of blood need and a lower level of COVID-19 infection risk related to blood donation mobilized experienced SWBDs to donate within 3 weeks. Intention-to-treat analyses and average-treatment-effect-on-the-treated estimations confirmed that Questionnaire 1 could motivate SWBDs to actually donate blood. CONCLUSION: Various measures could ease blood shortage during the COVID-19 epidemic. Administration of Questionnaire 1 could increase blood donations during the epidemic.

Effective methods for reactivating inactive blood donors: a stratified randomised controlled study.

BACKGROUND: Recruiting of sufficient numbers of donors of blood products is vital worldwide. In this study we assessed the efficacy and cost-effectiveness of telephone calls and SMS reminders for re-recruitment of inactive blood donors. METHODS: This single-centre, non-blinded, parallel randomised controlled trial in Guangzhou, China included 11,880 inactive blood donors whose last donation was between January 1 and June 30, 2014. The donors were randomly assigned to one of two intervention groups (telephone call or short message service [SMS] communications) or to a control group without intervention. SMS messages with altruistic appeal were adopted in the SMS group; in addition to altruistic appeal, reasons for deferral of blood donation were also asked in the telephone group. All participants were followed up for 1 year. The primary outcome was re-donation rate, and rates in different groups were compared by intention-to-treat (ITT) analysis and estimation of the average treatment effect on the treated (ATT). Secondary outcomes were the self-reported deterrents. Other outcomes included the re-donation interval, and the incremental cost-effectiveness ratio (ICER) of telephone calls and SMS reminders on re-recruitment. RESULTS: ITT analysis revealed no significant differences in the re-donation rate among the three groups. ATT estimations indicated that among compliers, telephone calls significantly increased re-donation compared to both SMS reminders and no intervention. Donor return behaviour was positively associated with receiving reminders successfully, being male, older age, and previous donation history. The SMS reminder prompted donors to return sooner than no reminder within 6 months, and according to ICER calculations, SMS reminders were more cost-effective than telephone calls. Donors reported time constraints as the most main causes of self-deferral in the telephone group, and altruistic appeal had a positive effect on these donors. CONCLUSIONS: Interventions to reactivate inactive blood donors can be effective, with telephone calls prompting more donors to return but at a greater cost than SMS messages. SMS reminder with altruistic appeal can urge donors to re-donate sooner within 6 months than no reminder. TRIAL REGISTRATION: NCT03366441 (Reactivation of Inactive Blood Donors). Retrospectively registered 4 December 2017.

CCL18 from tumor-associated macrophages promotes angiogenesis in breast cancer.

The infiltration of tumor-associated macrophages (TAMs) is associated with extensive angiogenesis, which contributes to a poor prognosis in breast cancer. However, anti-angiogenic therapy with VEGF-specific monotherapy has been unsuccessful in treating breast cancer, and the molecular mechanisms associated with chemoresistance remain unclear. Here, we investigated whether CCL18, a chemokine produced by TAMs, can stimulate angiogenesis in breast cancer, as well as the underlying mechanisms. Double immunohistochemical staining for CCL18 and CD34/CD31/vWF was performed in 80 breast cancer samples to study the correlation between CCL18+ TAMs and microvascular density (MVD). Cocultures of TAMs with human umbilical vein endothelial cells (HUVECs) were used to model the inflammatory microenvironment, and CCL18-induced angiogenesis was evaluated both in vitro and in vivo. We demonstrated that CCL18+ TAM infiltration positively associated with MVD in breast cancer samples, which was correlated with tumor metastasis and poor prognosis. We confirmed, both in vitro and in vivo, that CCL18 and VEGF synergistically promoted endothelial cell migration and angiogenesis. Conversely, blocking CCL18 or VEGF with neutralizing antibodies synergistically inhibited the promigratory effects of TAMs. Silencing PITPNM3, a putative CCL18 receptor, on the surface of HUVECs abrogated CCL18-mediated promigration and the enhancement of HUVEC tube formation, independently of VEGFR signaling. Moreover, CCL18 exposure induced the endothelial-mesenchymal transformation and activated ERK and Akt/GSK-3ß/Snail signaling in HUVECs, thereby contributing to its pro-angiogenic effects. In conclusion, our findings suggest that CCL18 released from TAMs promotes angiogenesis and tumor progression in breast cancer; thus, CCL18 may serve as a novel target for anti-angiogenic therapies.

[Human platelet lysates promotes the proliferation of mesenchymal stem cells in vitro].

OBJECTIVE: To investigate the effect of human platelet lysates (HPL) obtained from platelet-rich plasma on the proliferation and biological characteristics of human mesenchymal stem cells (MSCs) in vitro. METHODS: HPL was obtained by repeated freeze-thawing of human plateletes, and the MSCs separated by density gradient centrifugation from 6 donors were expanded in medium supplemented with 10% fetal bovine serum (FCS) or HPL at different concentrations. The optimal concentration of HPL for cells culture was determined according to the cell proliferation kinetics. The cultured MSCs were characterized for their proliferation, cell phenotype, and cell cycle distribution. RESULTS: The HPL-supplemented medium contained 4 essential growth factors for the growth of MSCs, namely platelet-derived growth factors (0.53∓0.06 ng/ml), basic fibroblast growth factor (37.5∓4.31 pg/ml), insulin-like growth factor-1 (0.15∓0.06 mg/ml) and transforming growth factor (5150∓463 pg/ml). Cultured in the presence of HPL at the optimal concentration of 7.5%, the MSCs displayed a spindle-shaped fibroblast-like morphology without obvious changes in the proliferation activity till passage 8 (P>0.05), similar to those of cells in FCS-supplemented culture medium. Flow cytometry and cell cycle analysis revealed no differences in the phenotypes or cell cycle distribution between the cells cultured in the presence of 7.5% HPL and 10% FCS. CONCLUSION: The culture medium supplemented by 7.5% HPL can promote the expansion of human MSCs and maintain the basic biological characteristics of the cells.

[Distribution of HCV genotypes in volunteer blood donors in Guangzhou].

OBJECTIVE: To determine the distribution of HCV genotypes among volunteer blood donors in Guangzhou. METHODS: Six-nine HCV RNA-positive samples were collected from volunteer blood donors in Guangzhou. NS5B fragments of HCV were amplified followed by DNA sequencing and phylogenetic analysis. RESULTS: HCV genotypes were determined for 67 samples. Among them, the subtypes 1b, 2a, 3a, 3b, 6a and 6n were detected at the frequencies of 37.31%, 4.48%, 7.46%, 4.48%, 44.78% and 1.49%, respectively. CONCLUSION: HCV 1b and 6a are the most predominant two subtypes among volunteer blood donors in Guangzhou.

[Study of the platelet GP specific antibodies and HLA antibodies expression in platelet transfusion refractoriness patients].

OBJECTIVE: To investigate the correlation between the platelet GP specific antibodies/HLA antibodies and platelet transfusion refractoriness (PTR). METHODS: Sixty-five patients with PTR were selected in this study and were genotyped for HLA-A and HLA-B as well as HPA systems by standard PCR-SSP assays. The platelet GP specific antibodies and HLA antibodies in serum and platelet elution were tested with a solid phase ELISA. RESULTS: The HLA-A/B antigens and the frequencies of HPA-1, 2, 4, 5, 6, 9, 15 antigens in PTR patients had no difference from those in healthy donors. The freguencies of HPA-3a and 3b were 0.65 and 0.35, respectively. There was statistical difference between the 65 PTR patients and the healthy donors in HPA-3 freguencies (P < 0.05). Twenty-four patients (36.9 %) only expressed HLA antibodies, and 14 (21.5%) expressed HLA and platelet GP specific antibodies. The highest expression of anti-HLA-A/B specific antibodies was -A*9(46.2 %)/-B*40(33.6%), respectively. In serum, GPIIb/IIIa was expressed (26.2%), followed by GPIa/IIa (21.5 %). In platelet elution, GPIIb/IIIa was expressed of 41.5% and GPIb/IX 41.5%. Pedigree study was carried out for 2 patients. The results showed that the platelet GP specific antibody/HLA antibody developed in PTR patients was highly related to the mismatch with the platelet antigen/HLA antigen in their parents. CONCLUSION: The expressions of the HLA and platelet GP specific antibodies are the most important reason in PTR, it's meaningful to explore the correlation between PTR and HLA and HLA-A/B antigen in guiding platelet transfusion.

Curcumin induces down-regulation of EZH2 expression through the MAPK pathway in MDA-MB-435 human breast cancer cells.

Curcumin, a natural compound isolated from turmeric, may inhibit cell proliferation in various tumor cells through a mechanism that is not fully understood. The enhancer of zeste homolog 2 (EZH2) gene is overexpressed in human breast cancers with poor prognosis. In this study, we observed a dose- and time-dependent down-regulation of expression of EZH2 by curcumin that correlates with decreased proliferation in the MDA-MB-435 breast cancer cell line. The curcumin treatment resulted in an accumulation of cells in the G(1) phase of the cell cycle. Further investigation revealed that curcumin-induced down-regulation of EZH2 through stimulation of three major members of the mitogen-activated protein kinase (MAPK) pathway: c-Jun NH2-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) and p38 kinase. These data suggest that an underlying mechanism of the MAPK pathway mediates the down-regulation of EZH2, thus contributing to the anti-proliferative effects of curcumin against breast cancer.

[Expression of anti-platelet glycoprotein specific antibodies and anti-HLA antibodies in idiopathic thrombocytopenic purpura].

In order to investigate the expression of the anti-platelet glycoprotein specific antibodies and anti-HLA antibodies in idiopathic thrombocytopenic purpura (ITP), 45 patients with ITP were selected in this study. An easy PCR-SSP assay was used to detect single-nucleotide polymorphisms or deletion in HPA and HLA systems. The anti-platelet glycoprotein specific antibodies and anti-HLA antibodies in plasma or platelet eluate were tested with a solid phase ELISA. The results indicated that the anti-platelet glycoprotein specific antibodies were detected in plasma or platelet eluate of 45 patients, among which anti-GPIIb/IIIa/and anti-GpIb/IX were most common. Both the anti-platelet glycoprotein specific antibodies and anti-HLA antibodies were found in plasma of 11 patients. Pedigree investigation in 2 patients (case 37 and case 40) was carried out, the results showed that anti-platelet glycoprotein specific antibodies and anti-HLA antibodies detected in 2 patients closely related to incompatibility with platelet antigens and HLA antigens in parents. In conclusion, the results suggested that detection of the anti-platelet glycoprotein specific antibodies and anti-HLA antibodies in plasma or platelet eluate in combination with investigation of clinical manifestation of patients is important for diagnosis of idiopathic thrombocytopenic purpura.

Reappraisal of peripheral NK cells in women with recurrent miscarriage.

The aim of this study was to examine the relationship between peripheral natural killer (NK) cells and recurrent miscarriage by improved methods. Peripheral NK cell measurement was carried out using flow cytometry of morning blood samples obtained in the early follicular phase, analysed within 8 h of collection. Eighty-five Chinese women with recurrent miscarriage who previously tested negative for autoantibodies, and 27 control subjects who were not using any hormonal methods for contraception, were recruited. No significant difference was found in the number of peripheral NK cells and their subsets between women with recurrent miscarriages and control subjects. Only 5% of women with recurrent miscarriage had high peripheral NK cells. The number of previous miscarriages did not appear to have an impact on the number of NK cells. In conclusion, there appears to be limited value in the routine measurement of peripheral NK cells in women with recurrent miscarriage.

[Biological appraisal of human bone marrow mesenchymal stem cells during ex-vivo expansion].

This study was aimed to investigate the characteristics of human bone marrow mesenchymal stem cells during ex-vivo expansion, MSCs were isolated from human bone marrow. At each passage, the characteristics of proliferation kinetics, osteogenic and adipogenic differentiation potential were analyzed, and cell morphology, surface markers were investigated as well. The karyotype analysis was done in different passage cells. The infection HIV, HCV, HBV and TP were detected by ELISA. Mycoplasma contamination in vitro was detected by PCR method. HLA-SBT was used to reanalyze the results of HLA antigens and alleles. STR genetic loci were detected by PCR in the MSC1, MSC2, MSC3 and MSC4. The results indicated that the proliferative ability and osteogenic potential decreased with the increase of passage number during culture expansion. The multiple differentiation potential of MSCs was maintained during their life span. Karyotype analysis showed that MSCs from 4 groups before passage 8 were normal. The expression of CD29, CD44, CD105, CD166 and CD73 were positive. The expression of CD14, CD34, CD45, CD80, CD86 were all negative. SBT was used to identify HLA-A, B, Cw, DRB1, DRPB1, DQ alleles in the MSC1, MSC2, MSC3, MSC4. The genetype of STR in the MSC1, MSC2, MSC3, MSC4 was different. MSC 3 was examined by TP-ELISA to confirm the infectious disease of TP. MSC2 was contaminated by mycoplasma at passage 5. It is concluded that culture expansion causes MSCs to gradually lose their stem cell properties. During ex-vivo expansion of MSCs, the osteogenic differentiation potential is decreased. MSCs before passage 8 can be a valuable subject for basic research and clinical application.

Overexpression of AIB1 in nasopharyngeal carcinomas correlates closely with advanced tumor stage.

AIB1, a candidate oncogene in breast cancer, is commonly amplified and overexpressed in several types of human cancers. In this study, expression and amplification of AIB1 in nasopharyngeal carcinoma (NPC) were studied by immunohistochemical analysis and fluorescence in situ hybridization using tissue microarrays, including 80 specimens of NPC and 20 specimens of nonneoplastic nasopharyngeal mucosa. In this NPC cohort, overexpression and amplification of AIB1 was detected in 36 (51%) of 71 and 3 (7%) of 46 NPCs, respectively. Overexpression of AIB1 was observed more frequently in NPCs in late T stages (T3/T4, 24/35 [69%]) than in earlier stages (T1/T2, 12/36 [33%]; P < .05). In addition, 18 (72%) of 25 NPCs with lymph node metastasis (N1-3) showed overexpression of AIB1; the frequency was significantly higher than that in NPCs without node metastasis (N0, 18/49 [39%]; P < .05). These findings suggest that overexpression of AIB1 in NPCs may be important in the acquisition of an invasive and/or metastatic phenotype.