Synthesis and Electrochemistry of LiNixMn2-xO4.-Samples of LiNixMn2-xO4 with 0 ¡ x ¡ 0.5 are prepared by solid state reaction of LiOH, Ni(NO3)2·6 H2O, and MnO2 at 750 • C as well as by a sol-gel method using Mn(OAc)2, Ni(NO3)2, and LiOH as precursors and carbon black as stabilizer (250-800 • C), The electrochemical behavior of the samples is studied in Li/LiNixMn2-xO4 coin-type cells. The potential vs. capacity curves show a plateau at 4. 1 V for the Li/LiMn2O4 cells. With increasing x, the length of this plateau decreases and a new plateau at 4.7 V appears. The shift of the plateau potential is caused by the higher binding energy (by 0.6 eV) of the Ni eg electrons compared to the Mn eg electrons. LiNi0.5Mn1.5O4 prepared by the sol-gel technique at a firing temp. of ≈600 . degree.C exhibits a capacity of ¿ 100 mAh/g at 4.7 V which is maintained over tens of cycles. -(ZHONG, Q.; BONAKDARPOUR, A.; ZHANG, M.; GAO, Y.; DAHN, J. R.; J. Electrochem.

The study was designed to investigate whether serum hepatitis B virus (HBV) RNA is a strong surrogate marker for intrahepatic HBV covalently closed circular DNA (cccDNA) compared with serum HBV DNA, hepatitis B surface antigen (HBsAg), and hepatitis B e antigen (HBeAg) in HBeAg-positive chronic hepatitis B (CHB) patients. Serum HBV RNA, HBV DNA, HBsAg, HBeAg, and intrahepatic cccDNA were quantitatively detected at baseline (n ϭ 82) and 96 weeks (n ϭ 62) after treatment with nucleos(t)ide analogue (NUC) in HBeAg-positive CHB patients. The correlations among serum HBV RNA, HBV DNA, HBsAg, HBeAg, and intrahepatic cccDNA levels were then statistically analyzed. The results showed that pretreatment intrahepatic cccDNA levels correlated better with serum HBV DNA levels (r ϭ 0.36, P Ͻ 0.01) than with serum HBV RNA levels (r ϭ 0.25, P ϭ 0.02), whereas no correlations were found between pretreatment intrahepatic cccDNA levels and HBsAg (r ϭ 0.15, P ϭ 0.17) or HBeAg (r ϭ 0.07, P ϭ 0.56) levels. At 96 weeks after NUC treatment, intrahepatic cccDNA levels correlated well with HBsAg levels (r ϭ 0.39, P Ͻ 0.01) but not with serum HBV RNA, HBV DNA, and HBeAg levels (all P Ͼ 0.05). Besides, the decline in the intrahepatic cccDNA level from baseline to week 96 correlated better with the reduction in the serum HBsAg levels than with the decreases in the levels of the other markers (for the HBsAg decline, r ϭ 0.38, P Ͻ 0.01; for the HBV DNA decline, r ϭ 0.35, P ϭ 0.01; for the HBV RNA decline, r ϭ 0.28, P Ͻ 0.05; for the HBeAg decline, r ϭ 0.18, P ϭ 0.19). In conclusion, the baseline serum HBV RNA level or its decline after 96 weeks of NUC therapy correlated with the corresponding intrahepatic cccDNA level, while it was less than that seen with serum HBV DNA at baseline and HBsAg (or its decline) at 96 weeks after treatment, respectively. KEYWORDS chronic hepatitis B, covalently closed circular DNA, HBV DNA, HBV RNA, hepatitis B surface antigen, nucleos(t)ide analogue H epatitis B virus (HBV) infection is a global health problem; an estimated 240 million persons are chronically infected, and about 650,000 people die annually due to chronic hepatitis B (CHB) worldwide (1). Though the currently available antiviral drugs can effectively reduce HBV DNA levels in serum of CHB patients, HBV may not be eliminated due to the persistence of HBV covalently closed circular DNA (cccDNA) in the infected hepatocytes, which represents the key HBV replicative intermediate (2). The fundamental role of intrahepatic cccDNA is as a template for transcription of all viral RNAs, including pregenomic RNA (pgRNA), which further produces the offspring virion

The purpose of this study was to explore the correlations of serum hepatitis B core-related antigen (HBcrAg) with intrahepatic Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) and HBV total DNA in hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) patients. Serum HBcrAg and other parameters, including HBV DNA, HBV RNA, HBeAg, hepatitis B surface antigen (HBsAg), hepatitis B core antibody (anti-HBc), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were quantitatively measured at baseline and follow-up time points. Intrahepatic HBV cccDNA and total DNA were quantitatively detected at baseline and 96 weeks. Grading of liver necroinflammation and staging of hepatic fibrosis were assessed at baseline and 96 weeks. Correlations between serum HBcrAg and other parameters were analyzed by Pearson’s correlation analysis. The results showed that pretreatment HBcrAg correlated significantly with HBV total DNA levels (r = 0.328, P = 0.003) in 82 CHB patients, and, after removing three outliers, with intrahepatic HBV cccDNA (r = 0.323, P = 0.004; n = 79). Serum HBcrAg correlated better with HBV cccDNA in patients with lower levels of serum HBV DNA (stratified by 7 log IU/ml of HBV DNA; r = 0.656, P = 0.003 versus r = −0.02, P = 0.866). Significant inverse correlations were found between HBcrAg and grade of liver necroinflammation (r = −0.245, P = 0.037), stage of hepatic fibrosis (r = −0.360, P = 0.002) at baseline. Serum HBcrAg presented significant correlation with intrahepatic HBV cccDNA in patients with HBeAg seroconversion at 96 weeks (r = 0.622, P = 0.006). The decrease in HBcrAg showed significant correlation with the decrease in HBV cccDNA after 96-week NA therapy (r = 0.282, P = 0.043). Serum HBcrAg also correlated significantly with other serum markers at baseline and 96 weeks of NA therapy. In conclusion, baseline HBcrAg and its decreased value were significantly correlated with the corresponding intrahepatic HBV cccDNA.

No abstract

The use of amnion and amniotic fluid (AF) are abundant sources of mesenchymal stem cells (MSCs) that can be harvested at low cost and do not pose ethical conflicts. In human and veterinary research, stem cells derived from these tissues are promising candidates for disease treatment, specifically for their plasticity, their reduced immunogenicity, and high anti-inflammatory potential. This work aimed to obtain and characterize bovine amniotic fluid mesenchymal stem cells (AFMSC). The bovine AF from the amniotic cavity of pregnant gilts in the early stages of gestation (3- and 4-m-old bovine embryos) was collected. AFMSCs exhibit a fibroblastic-like morphology only starting from the fourth passage, being heterogeneous during the primary culture. Immunofluorescence results showed that AFMSCs were positive for β-integrin, CD44, CD73 and CD166, but negative for CD34, CD45. Meanwhile, AFMSCs expressed ES cell markers, such as Oct4, and when appropriately induced, are capable of differentiating into ectodermal and mesodermal lineages. This study reinforces the emerging importance of these cells as ideal tools in veterinary medicine; future studies aimed at a deeper evaluation of their immunological properties will allow a better understanding of their role in cellular therapy.

Melatonin can modulate neural stem cell (NSC) functions such as proliferation and differentiation into NSC-derived pluripotent stem cells (N-iPS) in brain tissue, but the effect and mechanism underlying this are unclear. Thus, we studied how primary cultured bovine NSCs isolated from the retinal neural layer could transform into N-iPS cell. NSCs were exposed to 0.01, 0.1, 1, 10, or 100 μm melatonin, and cell viability studies indicated that 10 μm melatonin can significantly increase cell viability and promote cell proliferation in NSCs in vitro. Thus, 10 μm melatonin was used to study miR-302/367-mediated cell reprogramming of NSCs. We noted that this concentration of melatonin increased reprogramming efficiency of N-iPS cell generation from primary cultured bovine NSCs and that this was mediated by downregulation of apoptosis-related genes p53 and p21. Then, N-iPS cells were treated with 1, 10, 100, or 500 μm melatonin, and N-iPS (M-N-iPS) cell proliferation was measured. We noted that 100 μm melatonin increased proliferation of N-iPS cells via increased phosphorylation of intracellular ERK1/2 via activation of its pathway in M-N-iPS via melatonin receptors 1 (MT1). Finally, we verified that N-iPS cells and M-N-iPS cells are similar to typical embryonic stem cells including the expression of pluripotency markers (Oct4 and Nanog), the ability to form teratomas in vivo, and the capacity to differentiate into all three embryonic germ layers.

Although melatonin has been shown to exhibit a wide variety of biological functions, its effects on promoting differentiation of neural cells remain unknown. Wnt signaling mediates major developmental processes during embryogenesis and regulates maintenance, self-renewal, and differentiation of adult mammalian stem cells. However, the role of the noncanonical Wnt pathway during neurogenesis remains poorly understood. In this study, the amniotic epithelial cells ( AECs) were isolated from bovine amnion and incubated with various melatonin concentrations (0.01, 0.1, 1, 10, or 100 μm) and 5 × 10(-5) m all-trans retinoic acid (RA) for screening optimum culture medium of neural differentiation, compared with each groups, 1 μm melatonin and 5 × 10(-5) m RA were selected to induce neural differentiation of AECs, and then siMT1, siMT2, oWnt-4, and siWnt-4 were expressed in AECs to research role of these genes in neural differentiation. Efficiency of neural differentiation was evaluated after expressed above genes using flow cytometry. Cell function of neural cells was demonstrated in vivo using spinal cord injury model after cell transplantation, and damage repair of spinal cord was assessed using cell tracking and Basso, Beattie, Bresnahan Locomotor Rating Scale scores. Results demonstrated that melatonin stimulated melatonin receptor 1, which subsequently increased bovine amniotic epithelial cell vitality and promoted differentiation into neural cells. This took place through cooperation with Wnt-4. Additionally, following cotreatment with melatonin and Wnt-4, neurogenesis gene expression was significantly altered. Furthermore, single inhibition of melatonin receptor 1 or Wnt-4 expression decreased expression of neurogenesis-related genes, and bovine amniotic epithelial cell-derived neural cells were successfully colonized into injured spinal cord, which suggested participation in tissue repair.

Mesenchymal stem cells (MSCs) are multi-potential cells that are able to proliferate and differentiate into other cell types. Much research has been done on the MSCs from the umbilical cord (UCMSCs) in human, mice, and avian, but little literature has been published about these cells in big livestock. Here, we choose Luxi cattle as the experimental animal, we describe an external culture of the UCMSCs from it and summarize the biological characteristics of these cells, e.g., morphologic appearance, surface antigens, colony-forming ability, gene expression, and differentiation potential were detected via using immunofluorescence and reverse transcription polymerase chain reaction (RT-PCR). The induced cells, osteoblast, lipoblast, hepatocyte, islet cells, and neurocyte were identified by Alizarin red staining, Oil-red-O staining, Periodic acid-schiff staining, and Dithizone staining and RT-PCR detection for specific genes. Results suggest that biological characteristics of the UCMSCs were similar to those of MSCs previously analyzed. The primary UCMSCs were sub-cultured to passage 32, the UCMSCs expressed gene CD29, CD44, CD73, CD90, and CD166, induced cells illustrated typical staining, and expressed specific genes, which indicate that the UCMSCs could be a novel alternative source of MSCs for experimental and clinical applications.