Antifouling Gold-Inlaid BSA Coating for the Highly Efficient Capture of Circulating Tumor Cells.

Large amounts of coexisting contamination in complex biofluid samples impede the quantified veracity of biomarkers, which is the key problem for disease confirmation. Herein, amyloid-like transformed bovine serum albumin inlaid with gold nanoparticles was used as a coating (BGC) on a substrate composed of silicon nanowires (SW; BGC-SW) under ambient conditions. After modification with the recognition group, BGC-SW could serve as an outstanding platform for the selective separation and sensitive detection of biomarkers in complicated biosamples. First, the BGC on SW with a large surface area exhibits excellent adhesion resistance. The attached amounts of contaminations in biofluids were decreased by over 78% compared with native bovine serum albumin (BSA) as the blocking agent. This is because the phase-transformed BSA coating provides stronger interactions with the SW than bare BSA, which results in a tighter attachment and more uniform coverage of the BGC. Furthermore, the gold matrix laid inside the antiadhesive coating ensures simple cross-linking with the recognition groups to selectively capture various biomarkers in complex biofluids and create a gentle release method. Circulating tumor cells (CTCs) were chosen as template biomarkers to verify the application of A-BGC-SW (BGC-SW modified with sgc8-aptamer) in various separation processes of untreated biofluids. The results showed that approximately six cells could be captured from a 1 mL fresh blood sample containing only 10 CTCs. The easy fabrication and excellent antiadhesion property endow A-BGC-SW with great potential in the field of biological separation.

Determination of Breast Milk Cell Immune Function and Maternal Health Education.

It was to investigate the breast milk cell immune function and the effect of health education on pregnant and lying-in women. 100 primiparas were randomly divided into the control group (50 cases): routine health education; the test group (50 cases): prenatal breastfeeding health education based on the control group. Breastfeeding status, as well as breast milk immune cell composition at each stage, were compared between the two groups after intervention. After the intervention, the maternal feeding knowledge score of the test group (17.3 ± 2.4) points was significantly higher than that of the control group (14.1 ± 2.9) points (P < 0.05); the total feeding self-efficacy score of the test group was significantly higher than that of the control group at four weeks after delivery and eight weeks after delivery (P < 0.05); at eight weeks after delivery, 42 parturients in test group chose exclusive breastfeeding, significantly more than 22 parturients in the control group (P < 0.05); during colostrum, CD3+ accounted for (57.8 ± 4.2)%, CD4+ accounted for (31.5 ± 3.7)%, CD8+ accounted for (26.2 ± 2.4)%, CD4+/CD8+ was (1.2 ± 0.3), significantly higher than those of transitional milk and mature milk (P < 0.05); during colostrum, IFN-γ was (1.4 ± 0.4) µg/L, IL-8 was (1.4 ± 0.4) µg/L, significantly higher than those of mature milk (P < 0.05). Breast milk is beneficial to improve the immune function of newborns. It is necessary to perform health education for pregnant and lying-in women and improve the breastfeeding rate.

Thermostable properties of the equine infectious anemia virus nucleocapsid protein NCp11.

Retroviral nucleocapsid (NC) proteins are multifunctional nucleic acid binding proteins, playing critical roles in essentially every step of the viral replication cycle. As a small, basic protein, NC contains one or two highly conserved zinc-finger domains, each having an invariant CCHC motif, flanked by basic residues. In this study, we report for the first time, to our knowledge, the thermostable property of equine infectious anemia virus (EIAV) NCp11. About 43% of purified NCp11 remained soluble after incubation at 100 °C for 60 min, and heat-treated NCp11 maintained its abilities to bind to the E. coli RNA and the EIAV packaging signal sequence. At a very high degree of sequence occupancy, NCp11 inhibited first-strand cDNA synthesis catalyzed by either a commercial or the purified EIAV reverse transcriptase, and heat-treated NCp11 still inhibited the first-strand cDNA synthesis. We also found that protein concentrations, at a range from 0.1 to 0.9 µg/µl, have not affected the NCp11 thermostability significantly. However, NCp11 at acidic pH was more thermostable. Our findings highlight a new feature of the NC protein. Detailed understanding of NC's properties and functions will facilitate the development of effective and rational therapeutic strategies against retroviruses.

Carbon monoxide polyhemoglobin improves the therapeutic effect and relieves inflammation in the colon tissue of haemorrhagic shock/resuscitation rats.

OBJECTIVE: The objective of this study was to test the therapeutic effect of carbon monoxide polyhemoglobin (polyCOHb) in haemorrhagic shock/resuscitation and its underlying mechanisms. METHODS: 48 rats were divided into two experimental parts, and 36 rats in the first experiment and 12 rats in the second experiment. In the first experimental part, 36 animals were randomly assigned to the following groups: hydroxyethyl starch group (HES group, n = 12), polyhemoglobin group (polyHb group, n = 12), and carbon monoxide polyhemoglobin group (polyCOHb group, n = 12). In the second experimental part, 12 animals were randomly assigned to the following groups: polyHb group (n = 6), and polyCOHb group (n = 6). Then the anaesthetised rats were haemorrhaged by withdrawing 50% of the animal's blood volume (BV), and resuscitated to the same volume of the animal's withdrawing BV with HES, polyHb, polyCOHb. In the first experimental part, the 72h survival rates of each groups animals were measured. In the second experimental part, the rats' mean arterial pressure (MAP), heart rate (HR), blood gas levels and other indicators were dynamically monitored in baseline, haemorrhagic shock (HS), at 0point resuscitation (RS 0h) and after 1 h resuscitation (RS 1h). The concentrations of malondialdehyde (MDA), superoxide dismutase (SOD), tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) were measured by ELISA kits in both groups of rats at RS 1h. Changes in pathological sections were examined by haematoxylin-eosin (HE) staining. Nuclear factor erythroid 2-related factor 2 (Nrf2) and haem oxygenase-1 (HO-1) levels were detected by immunohistochemical analysis, while myeloperoxidase (MPO) levels were detected by immunofluorescence. DHE staining was used to determine reactive oxygen species (ROS) levels. RESULTS: The 72h survival rates of the polyHb and polyCOHb groups were 50.00% (6/12) and 58.33% (7/12) respectively, which were significantly higher than that of the 8.33% (1/12) in the HES group (p < 0.05). At RS 0h and RS 1h, the HbCO content of rats in the polyCOHb group (1.90 ± 0.21, 0.80 ± 0.21) g/L were higher than those in the polyHb group (0.40 ± 0.09, 0.50 ± 0.12)g/L (p < 0.05); At RS 1h, the MDA (41.47 ± 3.89 vs 34.17 ± 3.87 nmol/ml) in the plasma, Nrf2 and HO-1 content in the colon of rats in the polyCOHb group were lower than the polyHb group. And the SOD in the plasma (605.01 ± 24.46 vs 678.64 ± 36.37) U/mg and colon (115.72 ± 21.17 vs 156.70 ± 21.34) U/mg and the MPO content in the colon in the polyCOHb group were higher than the polyHb group (p < 0.05). CONCLUSIONS: In these haemorrhagic shock/resuscitation models, both polyCOHb and polyHb show similar therapeutic effects, and polyCOHb has more effective effects in maintaining MAP, correcting acidosis, reducing inflammatory responses than that in polyHb.

Preparation and exchange transfusion effect of a double polymerization human umbilical cord haemoglobin of red blood cell substitute.

The development of haemoglobin-based oxygen carrier (HBOC) is an excellent supplement to pre-hospital emergency blood transfusions. In this study, a new type of HBOC was prepared by using human cord haemoglobin (HCHb) and glutaraldehyde (GDA) and Bis(3,5-dibromosalicyl) fumarate (DBBF) to modify (DBBF-GDA-HCHb), the changes of physicochemical indexes during its preparation were evaluated, while a traditional type of GDA-HCHb was prepared, and the oxygen-carrying capacity of two type of HBOC was evaluated by a rat model of 135.0% exchange transfusion (ET). Eighteen SD male rats were selected, and were randomly divided into control group (5.0% albumin), DBBF-GDA-HCHb group and GDA-HCHb group. The 12 h survival rate of the C group was 16.67%, and the two HBOC groups were both 83.33%. Compared with GDA-HCHb, DBBF-GDA-HCHb can reduce lactic acid content by supplying oxygen to hypoxic tissues in a more timely manner, and can also can improve the reduction of MAP due to ischaemia.

Convenient exosome separation by phosphatidylserine targeting polymer brush materials.

We report phosphatidylserine targeting polymer brush materials to selectively separate exosomes. This method provides an efficient separation strategy with ordinary centrifuge force, which improves the integrity and purity of the exosomes. Compared with the common methods, the content of contaminated lipoprotein in the resulting exosomes decreased obviously.

Protective effect and mechanism of low P50 haemoglobin oxygen carrier on isolated rat heart.

The protection of the isolated heart is very important in heart transplantation surgery, meanwhile, the ischaemia/reperfusion (I/R) of the isolated heart is the main cause of its damage. A timely supply of oxygen can significantly improve the prevention of myocardial ischaemia, however, the cardioprotective solution does not have an oxygen supply function. Haemoglobin Based on Oxygen Carriers (HBOCs) is a kind of nano-oxygen drug, which can effectively and timely supply oxygen to hypoxic organs and tissues. However, the oxygen-carrying and releasing capacity (P50) is different with different HBOCs. The aim of our study was to investigate whether STS (a kind of cardioprotective solution, St Thomas Solution) +different P50 HBOCs provide superior myocardial protection and decrease myocardial injury compared to only STS in rats Langendorff isolated heart perfusion model. The results showed that STS + HBOCs can improve cardiac function at 37 °C for 35 min and 120 min, and reduce myocardial infarctions, pathological changes, and apoptosis of cardiomyocytes, and the STS + low P50 HBOCs is more effective than the other two higher P50 HBOCs. We further demonstrated the outstanding protective effect of STS + low P50 HBOCs on cardiac function, reducing myocardial infarctions and apoptosis of cardiomyocytes in rat Langendorff isolated heart perfusion model.

Alterations of intestinal microbiota in liver cirrhosis with muscle wasting.

OBJECTIVES: The intestinal microbiota plays an important role in the nutritional status and energy metabolism of the host. Liver cirrhosis is accompanied by muscle wasting or sarcopenia. The aim of this study was to to explore the changes in intestinal microbiota in patients with liver cirrhosis and muscle wasting by using metagenomics. METHODS: This was a cross-sectional study of patients with (n = 30) and without (n = 30) muscle wasting and age- and sex-matched healthy controls (n = 30) to evaluate changes in intestinal microbiota by metagenomic gene sequencing. Muscle wasting was determined by the third lumbar vertebrae skeletal muscle index (L3 SMI). RESULTS: The Shannon index, which represents species diversity, of patients in the muscle-wasting group (2.11 ± 0.88) was lower than in the non-muscle-wasting group (2.64 ± 0.68; P = 0.039), which was significantly lower than in the healthy control group (2.70 ± 0.53; P = 0.023). There were 17 microbial species with significant differences in relative abundance between the two groups (linear discriminant analysis score >2; P < 0.05). The relative abundance of Escherichia coli, Peptostreptococcus stomatis, and Bacteroides uniformis showed the most significant association with L3 SMI. CONCLUSIONS: There were compositional alterations in intestinal microbiota in patients with liver cirrhosis and muscle wasting. L3 SMI is closely related to E. coli, P. stomatis, and B. uniformis in liver cirrhosis. Further interventional studies are needed to confirm whether improving intestinal microbiota can improve the nutritional status of patients with liver cirrhosis.

Deficiency of apoptosis-stimulating protein 2 of p53 protects mice from acute hepatic injury induced by CCl4 via autophagy.

BACKGROUND: Apoptosis-stimulating protein 2 of p53 (ASPP2) has a variety of biological functions, and is involved in cellular apoptosis, autophagy and inflammatory reaction. However, the role of ASPP2 in acute hepatic injury remains unclear. METHODS: We established an animal model of acute hepatic injury by intraperitoneal injection of CCl4. The expression profile of ASPP2 was measured in wild type (ASPP2+/+) mice with acute hepatic injury induced by CCl4. Hepatic pathological changes and liver function, apoptosis, inflammation and autophagic levels were measured in ASPP2+/+and ASPP2 haploid deletion (ASPP2+/-) mice with acute hepatic injury, respectively. After 3-methyladenine (3-MA) treatment, indicators of hepatic injury were observed in ASPP2+/+and ASPP2+/- mice with CCl4 injection. RESULTS: During the development of acute hepatic injury, ASPP2 expression significantly upregulated at 24 h and 48 h after CCl4 injection. ASPP2 haplotype deletion protected against acute hepatic injury, and this was mainly reflected in decreased ALT and AST levels, less hepatic tissue hemorrhage and necrosis, and reduced cellular inflammation and apoptosis in ASPP2+/- mice compared with ASPP2+/+ mice with acute hepatic injury. ASPP2 haploid deletion activates autophagy in mice with acute hepatic injury, and protects mice from acute hepatic injury via the autophagic signal pathway. CONCLUSION: ASPP2 haplotype deletion protected mice against acute hepatic injury through autophagy activation, which inhibited inflammation and apoptosis in acute hepatic injury.

An optimal polymerization process for low mean molecular weight HBOC with lower dimer.

The new research tried to improve the distribution of molecular weight of Hb-based oxygen carriers (HBOC), a bottleneck of glutaraldehyde (GDA)-polymerization process. The orthogonal experiments were done on the basis of the early study of human placenta Hemoglobin (Hb)-crosslinked-GDA and three factors were selected including the molar ratio of GDA and Hb, Hb concentration, and the rate of the feeding GDA. The optimal match condition of polymerization process prepared for the purpose of lower mean molecular weight, content of super-weight molecule, and the content of dimer. The results showed that the molar ratio of GDA and Hb was the greatest influencing factor on the molecular weight distribution of polymerized-Hb, followed by the Hb concentration, and the last is the rate of feeding GDA. The optimum matching conditions had reached the objective that the mean molecular weight with 155.54 ± 5.79, the content of dimer with 17.23 ± 3.71, and content of super-weight molecule with 0.17 ± 0.09, and the results can be repeated in the 30 times expansion experiments.

Polymerized human placenta hemoglobin improves resuscitative efficacy of hydroxyethyl starch in a rat model of hemorrhagic shock.

This study was designed to investigate whether resuscitation with polymerized human placenta hemoglobin (PolyPHb) extended survival and the efficacy of hydroxyethyl starch (HES) in a rat model of hemorrhagic shock. After withdrawal of 39 ml of blood/kg, rats were randomly resuscitated with PolyPHb/HES or HES alone. Systemic, hemodynamic, and blood gas parameters, tissue oxygenation, and plasma erythropoietin (EPO) were evaluated. The results showed that resuscitation with PolyPHb/HES doubled the survival rate and maintained relatively high blood pressure and tissue oxygenation when compared with the results obtained using HES 130 alone. It is concluded that infusion of PolyPHb improves the resuscitative efficacy of HES in the present experimental conditions.