Evaluation of a rail logistics transmission system for the transportation of blood components within a medical centre.

BACKGROUND AND OBJECTIVES: Rail logistics transmission systems (RLTSs) are commonly used for the transportation of blood samples, pathological specimens and other medical materials in many hospitals, as they are rapid, secure, cost-effective and intelligent. However, few studies have evaluated blood component transportation from blood banks to the patient care areas of hospitals using RLTS. In this study, we evaluate the RLTS used for the transportation of blood components within a medical centre. MATERIALS AND METHODS: The dispatch of blood components, including packed red blood cells (pRBCs), fresh frozen plasma (FFP), cryoprecipitate and platelet units, from a blood bank to critical care areas or general wards was done using RLTS. Parameters such as the delivery time, temperature, physical integrity and blood component quality were evaluated via analytical testing using specimens obtained before and after transportation by RLTS. RESULTS: The turnaround time and temperature of all tested blood units via RLTS transportation were able to meet the clinical demands of blood component delivery (median time: 323 s [118-668 s]; temperature variation: 4.5-8.9°C for pRBCs and FFP and 21.5-23.5°C for cryoprecipitate and platelet units). Furthermore, parameters of pRBC quality, including the haemolysis index and potassium and lactate dehydrogenase levels in plasma, were not significantly different before and after transportation through RLTS. Similarly, RLTS transportation affected neither the basic coagulation test results in FFP and cryoprecipitate specimens nor platelet aggregation and activation markers in apheresis platelet specimens. CONCLUSION: Hospital-wide delivery of blood components via RLTS seems to be safe, reliable and cost-effective and does not have any negative impact on blood quality. Therefore, the establishment of standard criteria, protocols and guidelines based on further studies is needed.

Let-7c regulates proliferation and osteodifferentiation of human adipose-derived mesenchymal stem cells under oxidative stress by targeting SCD-1.

Osteoporosis is a progressive bone disease characterized by decreased bone mass and density, which usually parallels a reduced antioxidative capacity and increased reactive oxygen species formation. Adipose-derived mesenchymal stem cells (ADMSCs), a population of self-renewing multipotent cells, are a well-recognized source of potential bone precursors with significant clinical potential for tissue regeneration. We previously showed that overexpressing stearoyl-CoA desaturase 1 (SCD-1) promotes osteogenic differentiation of mesenchymal stem cells. Micro-RNAs (miRNAs) are noncoding RNAs recently recognized to play key roles in many developmental processes, and miRNA let-7c is downregulated during osteoinduction. We found that let-7c was upregulated in the serum of patients with postmenopausal osteoporosis compared with healthy controls. Levels of let-7c during osteogenic differentiation of ADMSCs were examined under oxidative stress in vitro and found to be upregulated. Overexpression of let-7c inhibited osteogenic differentiation, whereas inhibition of let-7c function promoted this process, evidenced by increased expression of osteoblast-specific genes, alkaline phosphatase activity, and matrix mineralization. The luciferase reporter assay was used to validate SCD-1 as a target of let-7c. Further experiments showed that silencing of SCD-1 significantly attenuated the effect of let-7c inhibitor on osteoblast markers, providing strong evidence that let-7c modulates osteogenic differentiation by targeting SCD-1. Inhibition of let-7c promoted the translocation of ß-catenin into nuclei, thus activating Wnt/ß-catenin signaling. Collectively, these data suggest that let-7c is induced under oxidative stress conditions and in osteoporosis, reducing SCD-1 protein levels, switching off Wnt/ß-catenin signaling, and inhibiting osteogenic differentiation. Thus, let-7c may be a potential therapeutic target in the treatment of osteoporosis and especially postmenopausal osteoporosis.

Overexpression of stearoyl-CoA desaturase 1 in bone marrow mesenchymal stem cells enhance the expression of induced endothelial cells.

BACKGROUND: Bone marrow mesenchymal stem cells (BM-MSCs) are capable of differentiating into endothelial cells in vitro and acquire major characteristics of mature endothelial-like expression of vWF and CD31. SFAs and lipid oxidation products have been linked with postprandial endothelial dysfunction. Consumption of SFAs impairs arterial endothelial function, while a Mediterranean-type MUFA-diet has a beneficial effect on endothelial function by producing a decrease in levels of vWF, TFPI and PAI-1. Stearoyl-CoA desaturase 1 (SCD1), which converts SFA to MUFA, is involved in the cellular biosynthesis of MUFAs from SFA substrates. High expression of SCD1 is corresponded with low rates of fatty acid oxidation, therefore it might reduce inflammatory responses and be beneficial for the growth of induced endothelial cells. Overexpression of SCD1 in BM-MSCs might increase the growth of induced endothelial cells. The goal of this research is to study the relationship between overexpression of SCD1 and the expression of induced endothelial cells in BM-MSCs in vitro. METHODS: The gene SCD1 was integrated into a lentiviral vector, and then 293 T cells were transfected by the connected product to produce a packaged virus. BM-MSCs were infected by the packaged virus. Cell culture and endothelial induction were performed. Fluorescent quantitative PCR of CD31, vWF and VE-cad was performed after 1 week and 2 weeks to test the growth of induced endothelial cells. RESULTS: The mRNA amount of CD31, vWF and VE-cad of the SCD1 overexpressed group was statistically higher than that of the empty vector (EV) group and that of the normal group after 1 week and 2 weeks, respectively (p < 0.05). Immunocytochemical staining of CD31 or vWF was detected by visualizing red color. CONCLUSIONS: This study suggested that overexpression of SCD1 in BM-MSCs could increase the expression of induced endothelial cells in vitro.

[SCD1 over-expression inhibits palmitic acid-induced apoptosis of rat BRL hepatocytes].

OBJECTIVE: To investigate the protective mechanism of stearoyl-CoA desaturase 1 (SCD1) over-expression against the pro-apoptotic affects of palmitic acid on hepatocytes using the rat BRL cell line. METHODS: Concentration effect curves were generated using the trypan blue exclusion test to assess the death rate of BRL cells upon exposure to a dilution series of palmitic acid. The multiplicity of infection (MOI) of a lentiviral expression vector, pGC-FU-GFP, was determined for the BRL cells. Unmanipulated BRL cells were divided into two groups: the non-palmitate groups were composed of ordinary cultured cells (CON) alone, infected with lentivirus empty expression vector (negative control, NC), and infected with lentivirus overexpressing SCD1 (SCD1-LV); the palmitate groups were composed of ordinary cultured cells plus palmitate (CON+) alone, infected with lentivirus empty expression vector plus palmitate (NC+), and infected with lentivirus overexpressing SCD1 plus palmitate (SCD1-LV+). SCD1 mRNA expression was detected by real-time PCR. Propidium iodide (PI) single-staining was used to detect apoptosis and assess the cell cycle. Inter-group differences were analyzed statistically. RESULTS: The death rate of BRL cells increased significantly after 72 h of exposure to 400 mumol/L palmitate (P less than 0.01). The MOI of pGC-FU-GFP in BRL cells was 20. The expression of SCD1 was significantly higher in the SCD1-LV and SCD1-LV+ groups than in the respective controls (vs. CON: F = 289, P less than 0.01; vs. CON+: F = 1522, P less than 0.01). Palmitate exposure led to decreased expression of SCD1 (CON+ vs. CON, F = 22, P less than 0.05 and NC+ vs. NC: F = 34, P less than 0.05). The ratio of S stage cells was similar in all non-palmitate groups (CON, NC and SCD1-LV, P = 0.137). However, there was a significant apoptotic peak and lower ratio of S stage cells in the control palmitate groups (CON+ and NC+) and the activity of cell proliferation was decreased as well. The ratio of apoptotic cells was decreased significantly in the SCD1-LV+ group compared to the CON+ group (P less than 0.01). CONCLUSION: The expression of SCD1 and its desaturation activity increased in BRL cells upon infection with the pGC-FU-SCD1-GFP lentiviral vector, suggesting that SCD1 over-expression can decrease palmitic acid-induced toxicity and apoptosis in hepatocytes.

Platelet PD-L1 inhibits storage-induced apoptosis by sustaining activation of the AKT signalling pathway.

Platelet apoptosis is irreversible under current storage conditions in blood banks. Studies have shown that programmed cell death ligand 1 (PD-L1) in tumour cells is required for neoplastic progression, tumour recurrence and metastasis by regulating apoptosis. However, whether PD-L1 is involved in storage-induced apoptosis in platelets remains poorly understood. In this study, we explored whether PD-L1 on platelets participated in the regulation of storage-induced apoptosis under blood bank conditions, as well as the underlying mechanism. Several apoptotic events in platelets from humans and PD-L1-knockout mice during storage under blood bank conditions were measured. The mechanism by which storage-induced apoptosis was regulated by platelet-intrinsic PD-L1 signalling was further investigated. Our results showed that PD-L1 in platelets progressively decreased. There was a strong negative correlation between platelet PD-L1 expression and the phosphatidylserine (PS) externalization rate and cleaved caspase-3 level and a positive correlation with anti-apoptosis protein Bcl-xl. Ex vivo, PD-L1-/- platelets stored at 22 °C showed rapid apoptosis via an intrinsic mitochondria-dependent pathway over time. Likewise, inhibiting PD-L1 signalling with BMS-1166 accelerated apoptosis by intrinsic mitochondria-dependent pathway. Coimmunoprecipitation analysis revealed that PD-L1 could bind AKT in platelets, and the binding capacity of both showed a progressive decrease with time. Finally, the decrease in PD-L1 expression levels during storage could be attributed to a complex process of progressive secretion. Therefore, platelet PD-L1 inhibits storage-induced apoptosis by sustaining activation of the AKT signalling pathway, which is expected to become a target for alleviating platelet storage lesions (PSLs) under current blood bank conditions.

Detection of Apolipoprotein E Gene Polymorphism and Blood Lipid Level in Hemodialysis Patients.

BACKGROUND: The aim was to investigate the clinical characteristics of lipid metabolism and the effect of apolipoprotein E (ApoE) gene polymorphism on lipid metabolism in hemodialysis patients. METHODS: The serum levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDLC), low-density lipoprotein cholesterol (LDLC), ApoA1, ApoB, ApoE and lipoprotein Lp(a) were detected by polymerase chain reaction-restriction fragment length (PCR-RFLP). RESULTS: The level of serum TG was significantly increased and the level of HDLC was significantly decreased in hemodialysis patients. Serum TG level was 33% higher than normal, and HDLC was 10.4% less than normal. The correlation analysis showed that TG level was significantly correlated with serum albumin level and extracorporeal circulation blood flow during dialysis. HDLC was significantly correlated with KT/V. The incidence of hypertension in hemodialysis patients was 73.6% and cardiovascular disease was 25%. The level of TG in the cardiovascular disease group was significantly higher than that in the non-cardiovascular disease group, and there was no significant difference between the hypertensive group and the non-hypertensive group. ApoE gene polymorphism test showed that the frequency of ApoE genotype ε3/3 and allele ε3 was the highest in hemodialysis patients, and the levels of TC, TG and LDLC were higher in ApoE genotype ε3/4 + ε4/4. CONCLUSION: The levels of serum TG and ApoB were significantly increased in patients with hemodialysis, and HDLC and other indexes were significantly decreased. The level of TG in patients with cardiovascular complications was significantly higher than in patients without complications. TG level was significantly correlated with serum albumin level and extracorporeal circulation blood flow during dialysis. HDLC was significantly correlated with KT/V. Hemodialysis patients who had ApoE allele ε4 are prone to lipid metabolism disorders.

[Effects of pentoxifylline on hepatic nuclear factor-kappa B signaling pathway and insulin resistance in nonalcoholic steatohepatitis rats induced by fat-rich diet].

OBJECTIVE: To explore the effects of pentoxifylline (PTX) on nuclear factor-kappa B (NF-kB) signaling pathway, insulin receptor substrates (IRSs) and glucose transporter 2 (GLUT2) expressions in livers in a rat model of nonalcoholic steatohepatitis (NASH). METHODS: Rats fed a fat-rich diet for 4 weeks were randomly allocated into two groups; the model group rats (n = 12) were fed a high-fat diet alone and the PTX group rats (n = 12) were fed a high-fat diet plus PTX (100 mg x kg(-1)/d(-1)) in drinking water. Meanwhile, rats (n = 6) fed a standard diet from the start served as controls. All the rats were sacrificed at the end of the 24th week. Hepatic NF-kappaB binding activity was measured by electrophoretic mobility shift assay (EMSA). The expression of tumor necrosis factor (TNF) alpha and inhibitor kappaB (IkappaBalpha) proteins in livers were determined by Western blot. Messenger RNA of IRS-1, IRS-2 and GLUT2 expressions were examined by RT-PCR. RESULTS: NF-kappaB binding activity was higher in the model group than that in the controls, while it was lower in the PTX group compared with that in the model group. The expression of TNFalpha protein was markedly increased in the model group (vs. the control group) but decreased in the PTX group (vs. the model group). The expression of IkappaBaalpha protein was decreased in the model group (vs. the control group) but increased in the PTX group (vs. the model group) to a certain extent. IRS-2 mRNA expression was markedly increased in the model group, and significantly decreased in the PTX group when compared with the model group (P less than 0.01). CONCLUSIONS: PTX could influence NF-kappaB signaling pathway and IRS expression in livers of NASH rats, which might be involved in the improvement of hepatic insulin resistance.

[A study of the changes of hepatic gene expression in the process of nonalcoholic fatty liver disease development using U230A oligonucleotide microarray].

OBJECTIVES: To explore the changes of hepatic gene expression during the course of nonalcoholic fatty liver disease (NAFLD) development in rats. METHODS: A rat model of NAFLD was developed by feeding the animals a high-fat diet for 24 weeks. Liver tissues of the model rats and the control rats were analyzed at different time points using rat U230A (Affymetrix GeneChip), which covers 15650 genes. RESULTS: Compared with the control rats, the number of genes expressed differently in the model group rats at 4 and 8 weeks was 426 and 540. The up-regulated genes among them were intracellular phosphorylase genes, metabolic enzyme genes, fatty acid binding protein genes, cytochrome P450 genes, cellular transcription and differentiation genes. The down-regulated genes were ionic channel genes, hormone receptor genes, and cytoskeleton genes. At the 12th week, the number of the genes expressed differently was 501, in which 352 were up-regulated genes, including genes related to inflammation and apoptosis such as interleukin and Toll-like receptor 4. At the 16th week, the number of the differently expressed genes was 665, with 430 up-regulated, such as those related to the inflammation and apoptosis genes and collagen I and fibrosis genes, however cell regeneration genes were down-regulated. At the 24th week the number was 663, of which fibroblast growth factor, transforming growth factor and insulin-like growth factor genes were up-regulated. Of all the differently expressed genes, the number of up-regulated genes was 128, including 10 lipogenic genes, 46 metabolic genes, 15 inflammation genes, 10 apoptosis genes, and 16 fibrosis genes; and the down-regulated genes were 52, including 6 hormone receptor genes, 5 cell regeneration genes and 11 electron transport genes. CONCLUSION: The changes of the hepatic gene expression of rats fed a fat-rich diet are related to the duration of the feeding, and are correlated with their histopathology in the livers.

The effects of intraoperative cryoprecipitate transfusion on acute renal failure following orthotropic liver transplantation.

PURPOSE: The definition of risk factors associated with acute renal failure (ARF) following orthotropic liver transplantation (OLT) is still controversial. Cryoprecipitate, which can supply fibrinogen and other coagulation factors, is widely used in OLT. However, the effects of intraoperative cryoprecipitate transfusion on ARF following OLT remain unclear. METHODS: In a series of 389 adult patients who received grafts from deceased donors and underwent their first OLT, the clinical correlation between intraoperative cryoprecipitate transfusion and ARF following OLT was retrospectively studied after adjusting for potential confounders. The distribution of ARF and the causes of death within the first year after OLT were also compared separately in patients with and without cryoprecipitate transfusion. RESULTS: The incidence of ARF in patients with cryoprecipitate transfusion was significantly higher than in patients without cryoprecipitate transfusion (15.9 vs. 7.8 %, p = 0.012). A nonlinear relationship between intraoperative cryoprecipitate transfusion and ARF following OLT was observed. The risk of ARF increased with the cryoprecipitate transfusion level up to the turning point (16 U) (adjusted OR 1.1, 95 % CI 1.1-1.2; p < 0.001). When the cryoprecipitate level exceeded 16 U, the level of cryoprecipitate transfusion was not associated with the risk of ARF (OR 0.95, 95 % CI 0.85-1.1; p = 0.319). Deaths within the first year after the operation occurred more frequently in cases with cryoprecipitate transfusion (22.9 vs. 14.2 %, p = 0.029). CONCLUSIONS: These findings suggested that intraoperative cryoprecipitate transfusion is associated with ARF following OLT. Cryoprecipitate transfusion during OLT should be performed carefully until more convincing evidence has been found.