Increased ZNF84 expression in cervical cancer.

PURPOSE: Little is known about ZNF84 gene. This study aims to investigate ZNF84 expression in cervical cancer (CC) and the effects of ZNF84 on CC. MATERIALS AND METHODS: Cervical cancer tissue specimens were collected from The First People's Hospital of Foshan. ZNF84 and Akt expression were detected by immunohistochemistry. The influence of ZNF84 on cell proliferation was detected by CCK-8 kits. The effects of ZNF84 on Akt protein and mRNA expression were detected by western blotting and qPCR, respectively. RESULTS: High expression of ZNF84 protein (80.0%) was detected within CC tissues while negative expression was found in normal cervical tissues. ZNF84 was specifically associated with tumor size (p = 0.018) and negatively associated with other indicators. Further, in squamous cell carcinoma, ZNF84 was associated with both TNM staging (p = 0.041) and tumor size (p = 0.041). In vitro, we used shZNF84 to inhibit the mRNA and protein expression of ZNF84, and showed marked inhibition of cancer cell proliferation by shZNF84. Furthermore, inhibition of ZNF84 down-regulated Akt. Ly294002 (an Akt inhibitor) decreased the cell inhibition ability of shZNF84, indicating the involvement of Akt. Finally, the relationship between ZNF84 and Akt in vivo showed positive correlation (p = 0.023). CONCLUSION: ZNF84 expression was increased in CC tissues and associated with tumor size. ZNF84 promoted cell proliferation which might involve Akt signal.

GO/iron series systems enhancing the pH shock resistance of anaerobic systems for sulfate-containing organic wastewater treatment.

In this study, the effect of pH shock during the treatment of sulfate-containing organic wastewater was investigated using an anaerobic fermentation system reinforced with graphene oxide (GO)/iron series systems. The results show that the anaerobic system with the GO/iron series systems exhibited enhanced resistance to pH shock. Among them, the GO/Fe0 system had the strongest resistance to pH shock, the systems of GO/Fe3O4 and GO/Fe2O3 followed close behind, while the blank system performed the worst. After pH shock, the CODCr removal rate, SO4 2- removal rate, and gas production of the GO/Fe0 group were significantly improved compared with those of the control group by 51.0%, 65.3%, and 34.6%, respectively, while the accumulation of propionic acid was the lowest. Further, detailed microbial characterization revealed that the introduction of the GO/iron series systems was beneficial to the formation of more stable anaerobic co-metabolic flora in the system, and the relative abundance of Geobacter, Clostridium, Desulfobulbus and Desulfovibrio increased after acidic and alkaline shock.

Decreased IL-37 expression in hepatocellular carcinoma tissues and liver cancer cell lines.

The role of IL-37 in cancer is currently largely unknown. The present study aimed to investigate IL-37 expression in hepatocellular carcinoma (HCC), paracancerous tissues (PT) and liver cancer cell lines, and their associations between IL-37 and NF-κB. A total of 65 HCC and 65 PT tissues were collected. The expression of IL-37 and NF-κB in tissues was detected by immunohistochemistry (IHC) and the data was analyzed using SPSS software. In the in vitro studies, IL-37 gene was transfected into HepG2 and MHCC97H cell lines with Lipofectamine 3000, and the protein regulation of NF-κB by IL-37 was verified by immunofluorescence (IF) and western blotting. In HCC, the positive expression rates of IL-37 and NF-kB were 21.5 and 95.4%, respectively. In PT, strong positive staining of IL-37and weak positive staining of NF-κB were observed. The normal expression levels of IL-37 and NF-κB, the increased IL-37 and decreased NF-κB induced by IL-37 gene transfection were observed through IF in cell lines. In terms of clinical significance, the difference in IL-37 expression between HCC and PT was statistically significant (χ2=55.05; P<0.001). IL-37 expression in HCC but not PT was negatively associated with serum AFP (χ2=6.522; P=0.039). IL-37 expression in PT was associated with sex (χ2=13.12; P=0.003) and tumor size (χ2=7.996; P=0.045). NF-κB expression in PT was associated with age, sex and BCLC stage. Notably, there was a negative correlation between IL-37 and NF-κB in HCC (r=-0.277; P=0.029) but not in PT (P>0.05). IL-37 overexpression downregulated the NF-κB protein by 56.50% in HepG2 cells (P<0.05) and 30.52% in MHCC97H cells (P<0.05). In conclusion, the expression of IL-37 in HCC and PT was specifically associated with serum AFP and tumor size, respectively. IL-37 expression was negatively correlated with NF-κB protein expression in HCC tissues and liver cancer cell lines.

LINGO-1 shRNA Loaded by Pluronic F-127 Promotes Functional Recovery After Ventral Root Avulsion.

Spinal root avulsion typically leads to massive motoneuron death and severe functional deficits of the target muscles. Multiple pathological factors such as severe neuron loss, induction of inhibitory molecules, and insufficient regeneration are responsible for the poor functional recovery. Leucine-rich repeat and immunoglobulin-like domain-containing Nogo receptor-interacting protein 1 (LINGO-1), a central nervous system (CNS)-specific transmembrane protein that is selectively expressed on neurons and oligodendrocytes, serves as a potent negative mediator of axonal regeneration and myelination in CNS injuries and diseases. Although accumulating evidence has demonstrated improvement in axonal regeneration and neurological functions by LINGO-1 antagonism in CNS damage, the possible effects of LINGO-1 in spinal root avulsion remain undiscovered. In this study, a LINGO-1 knockdown strategy using lentiviral vectors encoding LINGO-1 short hairpin interfering RNA (shRNA) delivered by the Pluronic F-127 (PF-127) hydrogel was described after brachial plexus avulsion (BPA). We provide evidence that following BPA and immediate reimplantation, transplantation of LINGO-1 shRNA lentiviral vectors encapsulated by PF-127 rescued the injured motoneurons, enhanced axonal outgrowth and myelination, rebuilt motor endplates, facilitated the reinnervation of terminal muscles, improved angiogenesis, and promoted recovery of avulsed forelimbs. Altogether, these data suggest that delivery of LINGO-1 shRNA by a gel scaffold is a potential therapeutic approach for root avulsion. Impact Statement In this study, we attempted transplantation of lentivirus (LV)/leucine-rich repeat and immunoglobulin-like domain-containing Nogo receptor-interacting protein 1 (LINGO-1)-short hairpin interfering RNA (shRNA) encapsulated by the Pluronic F-127 (PF-127) hydrogel into a brachial plexus avulsion (BPA)-reimplantation model. We found that administration of LV/LINGO-1 shRNA facilitates neuron survival and axonal regeneration, attenuates muscle atrophy and motor endplate (MEP) loss, enhances neovascularization, and promotes functional recovery in BPA rats. Co-transplantation of LV/LINGO-1 shRNA and gel reinforces the survival-promoting effect, axonal outgrowth, and angiogenesis in comparison with LV/LINGO-1 shRNA application alone. Our research provides evidence that LV /LINGO-1 shRNA delivered by PF-127 represents a new treatment strategy for BPA repair.

Embryonic stem cell-derived neural precursor cells improve memory dysfunction in Abeta(1-40) injured rats.

The versatility of neural precursor cells (NPCs) derived from mouse embryonic stem cells (ESCs) has recently rekindled interests in cell replacement strategies aimed at neurodegenerative diseases. We observed the survival, migration, differentiation and functional recovery of NPCs transplanted into the hippocampus of aggregated beta-amyloid (Abeta) peptide injured rats. Congo Red plaques, Fluro-jade B positive degenerating neurons and neuronal loss were observed in the Abeta-injured hippocampus of rats, accompanied with significant increases in escape latency and decrease in the ratio of exploratory time in a Morris water maze test. EGFP-expressing mouse ES cells were induced into Nestin-positive NPCs before transplantation into the Abeta-injured hippocampus. A marked decrease in escape latency and exploratory time were observed at least 16 weeks after transplantation compared to Abeta-injured animals without grafts. Grafted EGFP-expressing NPCs spread away from the injection tract and about 12.01+/-0.67% and 9.41+/-0.78% of NPCs differentiated into, respectively, GFAP- and NF200-positive cells 4 W after transplantation. These ratios gradually increased to 40.25+/-0.57% and 19.35+/-0.84% by 16 W. The restoration of hippocampal function by ESCs suggests that cell transplantation may be the effective choice to improve the cognitive function caused by Abeta injured.

[Neural-precursor-cells-phase is not the indispensable checkpoint of embryonic stem cell differentiation into insulin-producing cells].

Investigation on the pathways of embryonic stem cells differentiation into insulin-producing cells is of importance to pancreatic tissue-engineering. Instead of passing through the classic multi-step-inducing method, the expanded embryonic stem cells that were cultured and expanded in the presence of mouse embryonic fibroblast feed-layer and leukemia inhibitory factor (LIF) were induced into insulin-producing cells directly. The results showed a similar consequence from two different inducing cultures. Without passing through a so-called indispensable differentiation phase, the neural-precursor-cell-stage, the expanded embryonic stem cells could be induced into insulin-producing cells. The insulin-producing cells population resulting from our modified method were similar to that resulting from the classic multi-step method (passing through the neural-precursor-cells-stage), thus suggesting that neural-precursor-cell-phase is not the indispensable checkpoint of embryonic stem cell differentiation into insulin-producing cells. Embryonic stem cells can be induced into insulin-producing cell by classic multi-step inducing method or by direct inducing method.

Knockdown of p54nrb inhibits migration, invasion and TNF-α release of human acute monocytic leukemia THP1 cells.

54 kDa nuclear RNA- and DNA-binding protein (p54nrb) which is also called non-POU domain-containing octamer-binding protein (NONO) is known to be multifunctional involved in many nuclear processes. It was shown that p54nrb/NONO was closely related to the occurrence of erythroleukemia. Whether p54nrb/NONO plays a role in progress of human acute monocytic leukemia remains unknown. In the present study, we examined the effects of p54nrb/NONO silencing on the biological characteristics of human acute monocytic leukemia THP1 cells. The results showed that p54nrb was strongly expressed in THP1 cells, and knockdown of p54nrb slightly promoted proliferation and strongly inhibited motility and invasion of THP1 cells. Moreover, knockdown of p54nrb strongly decreased the release of TNF-α from THP1 cells by inhibiting certain process of TNF-α secretion, specially for the release of TNF-α induced by lipopolysaccharide (LPS). Notably, the infection of negative control shRNA-containing lentiviruses promoted the migration and the release of TNF-α induced by LPS in THP1 cells. All the above results demonstrated that p54nrb slightly inhibited THP1 cell proliferation, but significantly promoted migration, invasion and release of TNF-α induced by LPS in THP1 cells. The present study indicates that p54nrb is a powerful molecule involved in the regulation of cell motility and promotes the migration and invasion of THP1 cells, and it is more likely to be involved in the release of inflammatory mediators and the motility of inflammatory cells.

A modified method for generation of neural precursor cells from cultured mouse embryonic stem cells.

The pluripotency and high proliferative capacity of embryonic stem (ES) cells make them an attractive source of different cell types for biomedical research and cell replacement therapies. It has been demonstrated that ES cells can be induced into neural precursor cells (NPCs) under conditions. NPCs can be expanded in large numbers for significant periods of time to provide a reliable source of cells for transplantation in neurodegenerative disorders and injury of the central nervous system. This study describes a modified method for generation of NPCs from cultured mouse ES cells.

Critical protein GAPDH and its regulatory mechanisms in cancer cells.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), initially identified as a glycolytic enzyme and considered as a housekeeping gene, is widely used as an internal control in experiments on proteins, mRNA, and DNA. However, emerging evidence indicates that GAPDH is implicated in diverse functions independent of its role in energy metabolism; the expression status of GAPDH is also deregulated in various cancer cells. One of the most common effects of GAPDH is its inconsistent role in the determination of cancer cell fate. Furthermore, studies have described GAPDH as a regulator of cell death; other studies have suggested that GAPDH participates in tumor progression and serves as a new therapeutic target. However, related regulatory mechanisms of its numerous cellular functions and deregulated expression levels remain unclear. GAPDH is tightly regulated at transcriptional and posttranscriptional levels, which are involved in the regulation of diverse GAPDH functions. Several cancer-related factors, such as insulin, hypoxia inducible factor-1 (HIF-1), p53, nitric oxide (NO), and acetylated histone, not only modulate GAPDH gene expression but also affect protein functions via common pathways. Moreover, posttranslational modifications (PTMs) occurring in GAPDH in cancer cells result in new activities unrelated to the original glycolytic function of GAPDH. In this review, recent findings related to GAPDH transcriptional regulation and PTMs are summarized. Mechanisms and pathways involved in GAPDH regulation and its different roles in cancer cells are also described.

When aging-onset diabetes is coming across with Alzheimer disease: comparable pathogenesis and therapy.

Diabetes mellitus is a metabolic disorder that is characterized by high blood glucose because of the insulin-resistance and insulin-deficiency in Type 2, while the insulin deficiency due to destruction of islet cells in the pancreas in Type 1. The development of Type 2 diabetes is caused by a combination of lifestyle and genetic factors. Aging patients with diabetes are at increased risk of developing cognitive and memory dysfunctions, which is one of the significant symptoms of Alzheimer disease (AD). Also, over 2/3 of AD patients were clinically indentified with impairment of glucose. Cognitive dysfunction would be associated with poor self-care ability in diabetes patients. This review will briefly summarize the current knowledge of the pathogenesis of these two diseases and highlight similarities in their pathophysiologies. Furthermore, we will shortly discuss recent progress in the insulin-targeted strategy, aiming to explore the inner linkage between these two diseases in aging populations.

Targeted migration and differentiation of engrafted neural precursor cells in amyloid beta-treated hippocampus in rats.

OBJECTIVE: To observe the migration and differentiation of the neural precursor cells (NPCs) that derived from murine embryonic stem cells (ESCs) when they were transplanted into amyloid beta (A beta)-treated rat hippocampus. METHODS: MESPU35, a murine ESC cell line that express the enhanced green fluorescent protein (EGFP), was induced differentiation into nestin-positive NPCs by modified serum-free methods. The A beta plaques and the differentiation of the grafted cells were observed by immunofluorescent staining. RESULTS: Comparing 16 weeks with 4 weeks post-transplantation, the migration distance increased about 5 times; the rate of migratory NPCs differentiating into glial fibrillary acidic protein (GFAP)-positive cells kept rising from (30.41+/-1.45) % to (49.25+/-1.23) %, and the rate of NPCs differentiating into neurofilament 200 (NF200) positive cells increased from (16.68+/-0.95) % to (27.94+/-1.21) %. Meanwhile, the GFAP-positive cells targeting to the ipsilateral side of A beta plaques increased from 60.2% to 81.3%, while the NF200-positive cells increased from 61.3% to 84.1%. The migration distance had significant positive linear correlations to the neuronal differentiation rate (r = 0.991) and to the astrocytic differentiation rate (r = 0.953). CONCLUSION: Engrafted NPCs migrate targetedly to the A beta injection site and differentiate into neurons and astrocytes.