Impact of Platelet Hyperreactivity and Diabetes Mellitus on Ischemic Stroke Recurrence: A Single-Center Cohort Clinical Study.

Purpose: Ischemic stroke recurrence (ISR) is prevented by inhibiting platelet function. To investigate the impact of high on-treatment platelet reactivity (HTPR) assessed by thromboelastography (TEG) and its risk factors on ISR in individuals who have experienced acute ischemic stroke (AIS) receiving dual anti-platelet therapy (DAPT). Patients and Methods: At the end of follow-up, a total of 264 patients who met the criteria were enrolled in this cohort study. The primary endpoint event was a recurrence of ischemic stroke within 90 days of onset. Results: The ISR rate was 7.2% (19/264). The recurrence rate in the HTPR group was 15.1% (8/53), which was significantly higher than the 5.2% (11/211) in the non-HTPR group (p = 0.013), and the type 2 diabetes mellitus (T2DM) group (12.5%, 10/80) was also significantly higher compared to the non-T2DM group (4.9%, 9/184) (p = 0.028). T2DM was an isolated risk factor for HTPR (adjusted OR = 3.06, 95% CI 1.57-5.98, P = 0.001). Kaplan-Meier plots showed that the cumulative risk (CR) of ISR was statistically different in the HTPR and T2DM groups compared to the non-HTPR group (log-rank P = 0.009) and the non-T2DM group (log-rank P = 0.026), respectively. The HTPR and T2DM groups had greater hazard ratios (HR) of ISR than the non-HTPR (adjusted HR = 2.78, 95% CI 1.06-7.32, P = 0.038) and non-T2DM (adjusted HR = 2.64, 95% CI 1.01-6.92, P = 0.049) groups. Conclusion: Both HTPR and T2DM are linked to ISR. Platelet Inhibition Rate (PIR) of TEG can early identify patients who are at high risk for having another ischemic stroke in patients undergoing DAPT, and this study may offer more evidence in favor of clinically personalized treatment and secondary prevention tactics.

Egress Regulatory Factors: How Toxoplasma Exits from Infected Cells?

Toxoplasma gondii is an obligatory intracellular protozoan in the family Apicomplexa. It infects almost one-third of the world's population and causes toxoplasmosis, a prevalent disease. The parasite's egress from infected cells is a key step in the pathology caused by T. gondii. Moreover, T. gondii's continuous infection relies heavily on its capacity to migrate from one cell to another. Many pathways are involved in T. gondii egress. Individual routes may be modified to respond to various environmental stimuli, and many paths can converge. Regardless of the stimuli, the relevance of Ca2+ as a second messenger in transducing these signals, and the convergence of various signaling pathways in the control of motility and, ultimately, egress, is well recognized. This review attempts to outline intra- and extra-parasitic regulators that mediate T. gondii egress, and provides insight into potential clinical interventions and research.

Expression and Bioinformatics Analysis of Key miRNAs in Stored Red Blood Cells.

Introduction: Erythrocyte transfusion is the most common therapeutic procedure in hospitalized patients. Adding standard preservatives to red blood cells allows them to be stored for up to 42 days. However, whether storage has an effect on the erythrocyte transcriptome has not been well-studied. Objective: This study was designed to explore the change of key risk microRNA (miRNAs) in stored erythrocytes. Methods: We reanalyzed differentially expressed genes in the gene expression dataset GSE114990 and predicted their target genes, followed by experimental Gene Ontology (GO) analysis and (Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Furthermore, the PPI network of target genes was constructed by the STRING database, and the module analysis was carried out. Results: We found two differential miRNAs, which were hsa-miR-1245a and hsa-miR-381. Enrichment analysis of GO and KEGG pathways confirmed that these target genes were significantly enriched in organ and system development, anchoring junction, transcription factor binding, and pathways of cancer. Conclusion: The results suggest that the miRNAs hsa-miR-381 and hsa-miR-1245a may serve as biomarkers for storage products of erythrocytes.

microRNA-196a Overexpression Inhibits Apoptosis in Hemin-Induced K562 Cells.

microRNAs (miRNAs) have a crucial role in erythropoiesis. However, the understanding of the apoptosis of erythroid lineage remains poorly understood. Hence, an additional examination is required. K562 cell lines can be differentiated into early erythrocytes by hemin and the model of early erythrocytes can be established, consequently. miR-196a has been proven to take part in antiapoptosis in many cell lines. However, the role of miR-196a associated with the apoptosis in hemin-induced K562 cells remains unclear. To study the potential function of miR-196a involved in the common progenitor of erythroblasts, miR-196a mimics and microRNA-small hairpin negative control (miRNA-ShNC) were transfected into hemin-induced K562 cells with lentiviruses. After that, the viability of the transfected hemin-induced K562 cells was tested by CCK-8 assay, and the alteration of cell cycle and apoptosis rate were detected by flow cytometry. Furthermore, bioinformatics and dual-luciferase report system verified that p27kip1 is a target gene of miR-196a. Additionally, the expression of some proteins associated with cell cycle and apoptosis was tested by Western blotting assays. It was found that after overexpressing miR-196a, the proliferation of hemin-induced K562 cells was promoted while the apoptosis inhibited. Furthermore, miR-196a combines with the 3'UTR of p27kip1 directly. Additionally, the relationship between miR-196a and the protein level of p27kip1 is negative. After restoring the expression of p27kip1, the growth rate of hemin-induced K562 cells was not as high as before and the inhibition of apoptosis was alleviated. The present study validates that miR-196a overexpression inhibits apoptosis in hemin-induced K562 cells through downregulating p27kip1.

Bioinformatics analysis of the prognosis and biological significance of HMGB1, HMGB2, and HMGB3 in gastric cancer.

High mobility group box (HMGB) consists primarily of HMGB1, HMGB2, and HMGB3 proteins. Although abnormal HMGB expression is associated with various tumors, the relationship with gastric cancer (GC) remains unclear. In this study, HMGB1, HMGB2, and HMGB3 expression was analyzed using the Oncomine and TCGA databases. Correlations between HMGB1, HMGB2, and HMGB3 and clinicopathological factors were analyzed. cBioPortal was used to analyze HMGB1, HMGB2, and HMGB3 genetic alterations and its gene regulation network in GC tissue. HMGB1, HMGB2, and HMGB3 expression was higher in tumor tissues than in normal tissues, especially in GC. High HMGB1, HMGB2, and HMGB3 expression may predict a poor prognosis among patients with GC (hazard ratios [HR] = 1.90; 95% confidence interval [CI]: [1.30-2.78]) and human digestive system neoplasm (HR = 1.85; 95% CI [1.64-2.10]). These findings suggest that HMGB1, HMGB2, and HMGB3 may be useful prognostic indicators for patients with GC.

Curdione inhibits thrombin-induced platelet aggregation via regulating the AMP-activated protein kinase-vinculin/talin-integrin αIIbß3 sign pathway.

BACKGROUND: Curdione, a sesquiterpene compound isolated from the essential oil of Curcuma aromatica Salisb. inhibits platelet aggregation, suggesting its significant anticoagulant and antithrombotic effects. However, the mechanisms have not been fully elucidated. HYPOTHESIS: We hypothesized that curdione inhibits thrombin-induced platelet aggregation via regulating the AMP-activated protein kinase-vinculin/talin-integrin αIIbß3 signaling pathway. STUDY DESIGN: We performed in vitro assays to evaluate the effect of curdione on thrombin-induced expression levels of the AMPK signaling molecule and integrin αIIbß3 signaling pathway components. METHODS: Platelet proteins were extracted from washed human platelets, and the effects of curdione on thrombin-induced platelet aggregation were evaluated. The expression levels of the AMPK signaling molecule and integrin αIIbß3 signaling pathway-related proteins were examined using western blot and RT-PCR. The binding of vinculin and talin were studied using immunoprecipitation, double immunofluorescence staining and microscale thermophoresis. RESULTS: Platelet aggregation analysis showed that 0.02 U/ml thrombin significantly induces platelet aggregation. Western blot and RT-PCR analysis revealed that AMPK inhibits the vinculin/talin-mediated integrin αIIbß3 signaling pathway, and curdione downregulates the thrombin-induced expression of phosphorylated AMPK (P-AMPK) and P-integrin at both the protein and mRNA levels and downregulates vinculin and talin at the protein level. Furthermore, microscale thermophoresis experiments showed that curdione inhibits the binding of vinculin and talin. The results from the immunoprecipitation and double immunofluorescence staining were consistent with the results of the microscale thermophoresis experiments. CONCLUSION: Curdione inhibits thrombin-induced platelet aggregation via regulating the AMP-activated protein kinase-vinculin/talin-integrin αIIbß3 signaling pathway, which suggests its therapeutic potential in ethnomedicinal applications as an anti-platelet and anti-thrombotic compound to prevent thrombotic diseases.

Preventing murine transfusion-related acute lung injury by expansion of CD4+ CD25+ FoxP3+ Tregs using IL-2/anti-IL-2 complexes.

BACKGROUND: Transfusion-related acute lung injury (TRALI) is one of the most serious adverse events following transfusion, and there is no specific treatment in clinical practice. However, regulatory T cells (Tregs) have been suggested to play a potential role in the treatment of TRALI. This study investigated whether interleukin (IL)-2 or IL-2/anti-IL-2 complexes (IL-2c), which are mediators of Treg expansion, can modulate the severity of antibody-mediated TRALI in vivo. STUDY DESIGN AND METHODS: This study utilized a mouse model of the "two-hit" mechanism: BALB/c mice were primed with lipopolysaccharide (LPS) as the first hit, and then TRALI was induced by injecting major histocompatibility complex Class I antibodies. Mice injected with LPS only or LPS combined with isotype control antibodies served as controls. For the Treg-depleted groups, mice were infused with anti-mouse IL-2Rα first and then subjected to the same treatments as the TRALI group. Regarding IL-2- and IL-2c-treated mice, recombinant murine IL-2 or IL-2c was intraperitoneally administered to mice for 5 consecutive days before induction of the TRALI model. Samples were collected 2 hours after TRALI induction. RESULTS: Prophylactic administration of IL-2 or IL-2c to mice prevented the onset of edema, pulmonary protein levels, and proinflammatory factors that inhibited polymorphonuclear neutrophil aggregation in the lungs. Furthermore, the percentage of CD4+ CD25+ FoxP3+ Tregs was expanded in vivo using IL-2 and IL-2c compared to TRALI mice, as was confirmed through analysis of the spleen, blood, and lung. CONCLUSION: This study validates that the protective mechanisms against TRALI involve CD4+ CD25+ FoxP3+ Tregs, which can be expanded in vivo by IL-2 and IL-2c. This results in increased IL-10 levels and decreased IL-17A, thereby prophylactically preventing antibody-mediated murine TRALI.

Low STYK1 expression indicates poor prognosis in gastric cancer.

BACKGROUND: The expression of serine threonine tyrosine kinase 1 (STYK1), a member of the receptor protein tyrosine kinase (RPTK) family, is abnormal in several cancers. However, the molecular mechanism of STYK1 regulation of gastric cancer (GC) progression is unknown. MATERIALS AND METHODS: We evaluated STYK1 expression in GC tissues and the corresponding normal tissues. Specimens from 93 patients with GC were examined with immunohistochemical staining. The relationship between STYK1 protein expression and the patients' clinicopathological features was assessed. Kaplan-Meier and Cox proportional regression analyses were used to evaluate the association between STYK1 expression and survival. RESULTS: STYK1 expression was decreased in GC tissues. Low STYK1 expression was significantly associated with poor tumor differentiation (P=0.023), advanced clinical stage (P=0.021), and poor overall survival (OS; P=0.034). Univariate and multivariate analyses revealed that STYK1expression was an independent prognostic indicator (HR =0.53, 95% CI =0.29-0.95, P=0.039; HR =0.51, 95% CI =0.24-0.91, P=0.030, respectively). CONCLUSION: Downregulated STYK1 expression correlated significantly with poor tumor differentiation, advanced clinical stage, and poor OS in GC. STYK1 might be a diagnostic and prognostic indicator in patients with GC.

[Monitoring of Coagulation Status and Evaluation of Antiplatelet Aggregation in Patients with CHD by Using TEG].

OBJECTIVE: To evaluate the performance of thromboelastography (TEG) to monitor in vivo blood coagulation status and the efficacy of antiplatelet aggregation drugs in the patients with coronary heart disease (CHD) after oral anticoagulation. METHODS: Seventy one CHD patients were enrolled in CHD group and 380 healthy persons with normal TEG were enrolled in the control group. After admission, all CHD patients were administrated with routine anti-platelet aggregation drugs at a clinically recommended dose. Then, TEG was applied to monitor the basic blood coagulation indexes, such as R value, K value, α angle, MA value, CI value and a series of related indexes on platelet inhibition. RESULTS: Above 80% of the basic blood coagulation indexes in TEG were within normal reference range in the CHD group. the R value, MA value, α angle and CI value in the CHD group were not significanly different, from that in the control group, but the K value significantly increased (P<0.05). Compared with the control group, relatively higher ratio of male was included in the CHD patients at much older age (P<0.05), 83.1% of the CHD patients achieved significant anti-platelet aggregation effect (platelet inhibition rate>50%). Other antiplatelet aggregation indexes, MAADP, MAck and MAA suggested a 9.86%, 4.23% and 12.68% risk of thrombogenesis, respectively. Among all the related antiplatelet aggregation indexes, MAck showed the strongest correlation with age (correlation coefficient, 0.111), and ADP% most highly correlated with body mass (correlation coefficient, 0.160). CONCLUSION: TEG results can provide valuable coagulation information for clinicians, thus certainly guiding in the treatment for CHD patients receiving anti-platelet therapy. Moreover, the application of TEG can also provide accurate information for further individualized treatment of CHD patients, which would funther inprove the safety of anti-thrombotic therapy.

Inhibition of Suicidal Erythrocyte Death by Indirubin-3'-Monoxime.

BACKGROUND/AIMS: Qing Dai is a prized traditional Chinese medicine whose major component, indirubin, and its derivative, indirubin-3'-monoxime (IDM), have inhibitory effects on the growth of many human tumor cells and pronounced anti-leukemic activities. However, the effects of IDM on mature human erythrocytes are unclear. This study aimed to evaluate the potential impact of IDM on erythrocytes and the mechanisms underlying that impact. METHODS: Utilizing flow cytometry and confocal laser scanning microscopy, phosphatidylserine exposure at the cell surface was estimated by annexin V-fluorescein isothiocyanate (FITC). The relative cell size, expressed in arbitrary units, was evaluated by forward scatter in a flow cytometer. Fluo-3 fluorescence was used to bewrite changes in cytosolic Ca2+ activity, reactive oxygen species (ROS) formation was assessed by 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescence, and ceramide abundance was evaluated by FITC-conjugated specific antibodies. RESULTS: The 24-h exposure of human erythrocytes to IDM (12 µM) significantly decreased the percentage of annexin V-binding erythrocytes and the intracellular calcium concentration ([Ca2+]i). IDM (3-12 µM) did not significantly modify the ceramide level or DCFH-DA fluorescence. Energy depletion (removal of glucose for 24 hours) significantly increased annexin V binding and Fluo-3 fluorescence and diminished forward scatter, and these effects were significantly mitigated by IDM (12 µM). Moreover, the Ca2+ ionophore ionomycin (1 µM, 60 min) and oxidative stress (30 min exposure to 0.05 mM tert-butyl hydroperoxide, t-BHP) similarly triggered eryptosis, which was also significantly suppressed by IDM. CONCLUSIONS: IDM is a novel inhibitor of suicidal erythrocyte death following ionomycin treatment, t-BHP treatment and energy depletion. Thus, IDM may counteract anemia and impairment of microcirculation, at least in part, by inhibition of Ca2+ entry into erythrocytes.

Microparticles in red cell concentrates prime polymorphonuclear neutrophils and cause acute lung injury in a two-event mouse model.

Red cell-derived microparticles (RMPs) are potential mediators of transfusion-related acute lung injury (TRALI). The aim of this study was to investigate the effects of microparticles present in red cell concentrates (RCC) on polymorphonuclear neutrophil (PMN) respiratory burst and acute lung injury (ALI) in mice. Microparticles (MPs) in RCC supernatant were quantified using flow cytometry. The priming activity of either isolated MPs or RCC supernatant toward human PMN was measured in vitro. Mice were injected with lipopolysaccharide (LPS), followed by an infusion of either isolated MPs or heat-treated RCC supernatant. The lungs were harvested to assess myeloperoxidase (MPO) activity, histology and pulmonary edema. Protein content in bronchoalveolar lavage fluid (BALF) was measured. The number of RMPs increased significantly during storage. Both isolated MPs and the supernatants from RCCs that had been stored for 28 and 35days effectively primed the PMN respiratory burst. The infusion of isolated MPs or supernatants that had been stored for >28days into LPS-treated mice caused ALI. The filtered supernatant resulted in significantly ameliorated mouse ALI. MPs that accumulate during RCC storage prime the PMN respiratory burst and cause ALI in a two-event mouse model.

Sequential Growth of NaYF4:Yb/Er@NaGdF4 Nanodumbbells for Dual-Modality Fluorescence and Magnetic Resonance Imaging.

Upconversional core-shell nanostructures have gained considerable attention due to their distinct enhanced fluorescence efficiency, multifunctionality, and specific applications. Recently, we have developed a sequential growth process to fabricate unique upconversion core-shell nanoparticles. Time evolution of morphology for the NaYF4:Yb/Er@NaGdF4 nanodumbbells has been extensively investigated. An Ostwald ripening growth mechanism has been proposed to illustrate the formation of NaYF4:Yb/Er@NaGdF4 nanodumbbells. The hydrophilic NaYF4:Yb/Er@NaGdF4 core-shell nanodumbbells exhibited strong upconversion fluorescence and showed higher magnetic resonance longitudinal relaxivity (r1 = 7.81 mM-1 s-1) than commercial contrast agents (Gd-DTPA). NaYF4:Yb/Er@NaGdF4 nanodumbbells can serve as good candidates for high efficiency fluorescence and magnetic resonance imaging.

Reference values for kaolin-activated thromboelastography in volunteers of Anhui Province in China.

BACKGROUND: The assessment of the coagulation status using thromboelastography (TEG) in Chinese population has less been reported. This study aimed to establish reliable reference values for kaolin-activated TEG in Chinese volunteers. METHODS: A total of 1681 Chinese adult individuals were recruited for this study. The reference individuals were stratified by gender and age, and the TEG values were measured on the basis of strict quality control. The 95% reference values were determined using nonparametric statistical methods. RESULTS: The sex-related 95% reference values were reaction time (R):4.2-8.7 minutes; clotting time (K): 1.2-3.2 minutes; alpha angle (α): 47.0-72.3 degree; maximum amplitude (MA): 49.1-70.5 mm for males, and R: 3.7-9.0 minutes; K: 1.0-3.2 minutes; α: 48.4-74.4 degree; MA: 46.8-72.4 mm for females. Also, the TEG parameters indicated a relatively more hypercoagulable profile in both female and elder groups. CONCLUSIONS: This study established the reference values for kaolin-activated TEG in the target Chinese population, which might provide a reference for both clinical and laboratory studies.

GADD45α is involved in the apoptosis of lymphocytes induced by riboflavin and ultraviolet light.

BACKGROUND: Riboflavin plus ultraviolet (UV) pathogen reduction technology (RF-PRT) is an effective method for inactivating the residual white blood cells (WBCs) in blood components. The RF-PRT system for platelets is known to activate many signaling pathways, including p38 and NF-κB. Nevertheless, proteomic studies in WBCs after riboflavin plus UV treatment requires further analysis. STUDY DESIGN AND METHODS: ABO/D-matched lymphocytes were pooled, split, and treated with RF-PRT or UV light or left untreated. After treatment, cell apoptosis was measured. In addition, cell proliferation and the cycle distribution were evaluated upon stimulation with phytohemagglutinin. The changes in the protein expression levels of growth arrest and DNA damage-inducible (GADD)45α, p38, and c-Jun N-terminal kinase (JNK) were determined by Western blotting. The effect of GADD45α, p38, and JNK on apoptosis was assessed. RESULTS: RF-PRT significantly inhibited proliferation and induced G1 arrest in lymphocytes. Furthermore, the percentage of apoptotic cells was increased in RF-PRT-treated lymphocytes compared to UV-treated cells or untreated cells, associated with the up regulation of GADD45α expression. Consistent with these observations, the inhibition of GADD45α expression partially counteracted the effects of riboflavin plus UV treatment. The p38 and JNK signaling pathways were activated by GADD45α in RF-PRT-treated lymphocytes. CONCLUSIONS: These data revealed that RF-PRT effectively inhibited proliferation and induced apoptosis of lymphocytes by promoting GADD45α expression, which subsequently activates p38 and JNK signaling pathways.

Eryptosis as an Underlying Mechanism in Systemic Lupus Erythematosus-Related Anemia.

BACKGROUND: The progression of systemic lupus erythematosus (SLE) leads to anemia in patients, adversely affecting prognosis. The diverse causes of anemia may include excessive eryptosis or premature suicidal erythrocyte death characterized by cell shrinkage and phosphatidylserine (PS) exposure on the cell surface. The present study explored if SLE enhances eryptosis and the underlying mechanisms. MATERIALS AND METHODS: Eryptosis was assessed using flow cytometry in healthy volunteers (n = 20) and anemic patients hospitalized for SLE (n = 22), for parameters including PS exposure, cell volume, cytosolic calcium ion (Ca2+) levels and reactive oxygen species (ROS) and ceramide abundance. These indicators were measured in erythrocytes of experimental subjects and erythrocytes treated with plasma from healthy volunteers or SLE patients. RESULTS: The hemoglobin and hematocrit levels were significantly lower in anemic SLE patients than in healthy volunteers (***p<0.001, p<0.001, respectively). The percentage of PS-exposing erythrocytes was significantly higher in SLE patients than in healthy volunteers (p<0.001), accompanied by an increase in cytosolic Ca2+ levels, oxidative stress. The measurements of PS and Ca2+ levels were significantly higher in the erythrocytes of healthy volunteers following incubation in plasma of SLE patients than in plasma of healthy volunteers for 24h (***p<0.001, *p<0.05 respectively). CONCLUSION: Eryptosis is enhanced in SLE and may contribute to anemia. The probable underlying mechanisms may be an excessive formation of ROS in erythrocytes. Also, some plasma components may trigger eryptosis by increasing the cytosolic Ca2+ concentration.

A boswellic acid-containing extract ameliorates schistosomiasis liver granuloma and fibrosis through regulating NF-κB signaling in mice.

Boswellic acid (BA)-containing extracts such as BSE have anti-inflammatory and immunomodulatory activity. In chronic schistosomiasis, the hepatic granuloma and fibrosis induced by egg deposition in the liver is the most serious pathological manifestations. However, little is known regarding the role of BAs in Schistosoma japonicum (S. japonicum) egg-induced liver granuloma and fibrosis. In order to investigate the effect of a water-soluble complex preparation of BSE, BSE-CD, on S. japonicum egg-induced liver pathology, liver granuloma and fibrosis were induced by infecting C57BL/6 mice with 18-22 cercariae of S. japonicum. S. japonicum cercariae infected mice were injected with BSE-CD at the onset of egg granuloma formation (early phase BSE-CD treatment after 4 weeks infection) or after the formation of liver fibrosis (late phase BSE-CD treatment after 7 weeks infection). Our data show that treatment of infected mice with BSE-CD significantly reduced both the extent of hepatic granuloma and fibrosis. Consistent with an inhibition of NF-κB signaling as evidenced by reduced IκB kinase (IKK) activation, the mRNA expression of VEGF (vascular endothelial growth factor, VEGF), TNF-α (tumor necrosis factor-alpha TNF-α) and MCP-1 (monocyte chemotactic protein 1, MCP-1) was decreased. Moreover, immunohistochemical analysis (IHC) revealed that the content of α-SMA in liver tissue of BSE-CD treated mice was dramatically decreased. Our findings suggest that BSE-CD treatment attenuates S. japonicum egg-induced hepatic granulomas and fibrosis, at least partly due to reduced NF-κB signaling and the subsequently decreased expression of VEGF, TNF-α, and MCP-1. Suppression of the activation of hepatic stellate cells (HSC) may also be involved in the therapeutic efficacy of BSE-CD.

[Peripheral blood stem cell collection from ABO incompatible allogeneic donors and its effect in hematopoietic stem cell transplantation].

The study was aimed to evaluate the yield of the COBE Spectra blood cell separator with auto-peripheral blood stem cell program for collection of peripheral blood hematopoietic stem cells (PBHSC) from HLA-matched ABO-incompatible allogeneic PBHSC donor, and observe the safety and effect of allogeneic peripheral blood hematopoietic stem cell transplantation (allo-PBHSCT) without removal of erythrocytes and plasma. PBHSC from 28 allogeneic donors were collected by COBE Spectra blood cell separator with auto-peripheral blood hematopoiEtic stem cell (auto-PBHSCT) program. Control group included 15 HLA-matched patients who received allo-PBHSCT with ABO-compatible grafts. The amount of PBHSC was harvested and the parameter was modified according to the hematocrit and mononuclear cell (MNC) counts of donors. The nucleated cell count, proportion of MNC, number of CD34(+) cells were detected, and reconstitution status of hematopoietic function and time for change into donor's blood group were observed. The results showed that the nucleated cell count proportion of MNC and number of CD34(+) cells showed no significant difference between groups of ABO incompatible and compatible (p > 0.05). All their hematopoietic functions were reconstituted. Between the ABO incompatibility and the compatible groups, the time of neutrophil and platelet recovery was not significantly different (p > 0.05), In ABO blood major incompatible and the compatible groups, the recovery of erythropoiesis were significantly delayed (p < 0.01). The blood type of 18 patients in ABO incompatible group was turned into donor's blood type successfully at 35-139 days after transplantation. It is concluded that major ABO incompatibility did not affect the erythropoiesis reconstitution in HLA matched allo-HSCT. the major incompatibility may be a main reason of erythropoietic delay.

Application of signaling protein 14-3-3 and 26 kDa glutathione-S-transferase to serological diagnosis of Schistosomiasis japonica.

Schistosomiasis japonica is currently one of the most serious parasitic diseases and over 670000 people are infected in China by the end of 2006. In order to establish an effective diagnostic method, the gene coding for Sj14-3-3 and Sj26kDa GST were cloned and expressed separately in Escherichia coli as fusion protein with His-tag. The rSj14-3-3 and 26kDa rSjGST were combinedly used as antigens for enzyme-linked immunosorbent assays (ELISA) to diagnose acute and chronic S. japonica. Our results showed that the sensitivity in diagnoses of both acute and chronic schistosomiasis was 94.4% (67/71) and 80.7% (96/119), respectively. The specificity was 94.7% applying 132 sera from people living in S. japonicum-free areas. The data also showed that the recombinant proteins cross-react with Clonorchis sinensis and hookworms at a rate of 11.8% and 5.3% respectively. Parallel tests were conducted among ELISA, indirect hemagglutination assay (IHA) and circular ovum precipitin test (COPT) to determine anti-S. japonicum antibodies in sera of patients with schistosomiasis, healthy control, and those infected with other parasites and the results showed no significant difference in sensitivity for acute schistosomiasis between ELISA and IHA assays (chi(2)=1.33, P>0.05), but significant between ELISA and COPT assays (chi(2)=6.72, P<0.01). Our results also revealed significant difference in positive rate between ELISA and IHA (chi(2)=24.74, P<0.005), as well as between ELISA and COPT (chi(2)=58.14, P<0.005). These results suggest that the rSj14-3-3 and r26kDa SjGST would be effective diagnostic antigens for detection of antibodies to S. japonicum in human. Due to the easy production, high sensitivity and specificity, the recombinant proteins tested in this study can be considered as candidate reagent for immunological diagnosis of human schistosomiasis.