Anti-tumor effect of single-chain antibody to Reg3a in colorectal cancer.

BACKGROUND: Regenerating protein 3a (Reg3a) is a trophic factor that functions as a stimulus in cell proliferation and neogenesis. Previous studies showed that Reg3a is ectopically upregulated in a majority of colorectal cancers (CRC) and detectable in the serum. METHODS: Single-chain variable fragment targeting Reg3a (scFv-Reg3a) was screened from a phage library. The bioactivity of recombinant Reg3a (rReg3a) and scFv-Reg3a were tested in LoVo and RKO cell lines using MTT, flow cytometry, wound healing and transwell analyses. Whether scFv-Reg3a inhibits tumor growth and enhances 5-fluorouracil (5-FU)-caused cell death were further examined in LoVo cell-transplanted nude BALB/c mice. RESULTS: A scFv-Reg3a from clone C2 was obtained and its binding affinity (KD) to rReg3a was determined to be 4.44 × 10-10. In cultured LoVo and RKO cells, rReg3a promoted but scFv-Reg3a inhibited cell proliferation, survival, migration and invasion. In LoVo cell-xenografted nude mice, administration of rReg3a accelerated tumor growth while scFv-Reg3a suppressed cell proliferation and reinforced 5-FU-induced cell death. CONCLUSION: The newly developed scFv-Reg3a is an anti-cancer agent which is potent to suppress CRC cell proliferation and survival. The use of scFv-Reg3a could enhance the effectiveness of 5-FU-based chemotherapy in the cancerous treatment.

Dimorphic autoantigenic and protective effects of Reg2 peptide in the treatment of diabetic ß-cell loss.

AIMS: The potential effect of regenerating (Reg) proteins in the treatment of diabetes has been indicated in the past decade, but the clinical use of Reg proteins requires more advances in translational medicine. In the present study, we produced recombinant regenerating protein 2 (rReg2), to prove its protective effect against streptozocin (STZ)-induced diabetes in BALB/c mice. MATERIALS AND METHODS: rReg2 was administrated in STZ-induced diabetic mice. Blood glucose, body weight, serum insulin and islet ß-cell loss were determined. However, Reg2 has also been reported to serve as an autoantigen that induces autoimmune attacks on islets and aggravates diabetic development in non-obese diabetic mice. To address this contradiction, complete Freund's adjuvant was injected to generate a model that was hypersensitive to Reg2. In this model, islet CD8 T-cell infiltration, serum Reg2 antibody and interleukin (IL)-4 and IL-10, and splenic CD4+/interferon (IFN)-γ+ T cells were determined. RESULTS: Direct rReg2 pretreatment preserved islet ß-cell mass against STZ and improved glycaemia, body weight and serum insulin content. The protection against cell death was further confirmed in cultured mouse islets and MIN6 cells. On the other hand, significant elevations of serum Reg2 antibody and splenic CD4+/IFN-γ+ T cells, and decreases in serum IL-4 and IL-10 were detected in rReg2-vaccinated mice, which may contribute to the accelerated diabetes. Interestingly, these mice, upon further rReg2 treatment, exhibited alleviated diabetic conditions with less islet CD8+ T-cell infiltration. CONCLUSION: rReg2 treatment ameliorated STZ-induced diabetes in normal BALB/c mice. By contrast, rReg2 vaccination exacerbated, but further rReg2 treatment alleviated, the severity of STZ-induced diabetes. Thus, the protective effect of rReg2 is predominant over the autoantigenic ß-cell destruction, supporting the potential of rReg2 in the clinical treatment of diabetes.

Preparation of green high-performance biomass-derived hard carbon materials from bamboo powder waste.

Efficient energy storage systems are crucial for the optimal utilization of renewable energy. Sodium-ion batteries (SIBs) are considered potential substitutes for next-generation low-cost energy storage systems due to the low cost and abundance of sodium resources. However, the industrialization of SIBs faces a great challenge in terms of the anode. Hard carbon could be a promising anode material due to its high capacity and low cost which originates from biomass. This study used pre-treatment and template carbonization methods to extract a hard carbon material from a large amount of discarded biomass in bamboo powder waste. This material has a good initial Coulombic efficiency of 78.6 % and good cycling stability when applied to sodium ion batteries.Typically, the optimal hard carbon material is used as the anode to prepare sodium ion battery prototypes to demonstrate their potential applications. The anode exhibited excellent sodium storage performance with a reversible capacity of 303 mAh ⋅ g-1 at 1 C rate and good cycling performance, retaining 92.0 % of its capacity after 100 cycles. These results demonstrate that BPPHC is a promising candidate for anode material in sodium-ion batteries. This work suggests that bamboo powder could be a low-cost anode material for SIBs.

Single-chain Antibody Against Reg4 Suppresses Gastric Cancer Cell Growth and Enhances 5-FU-induced Cell Death in vitro.

BACKGROUND: Regenerating islet-derived gene family member 4 (Reg4), a well-investigated growth factor in the regenerative pancreas, has recently been reported to be highly associated with a majority of gastrointestinal cancers. Pathological hyper-expression or artificial over-expression of Reg4 causes acceleration of tumor growth, migration, and resistance to chemotherapeutic 5-Fluorouracil (5-FU). Until now, no method has been successfully established for eliminating the effects of Reg4 protein. METHODS: This study reports the production of an engineered immunoglobin, a single-chain variable fragment (scFv-Reg4), to specifically bind Reg4 and block the bioactivity. The complementary-determining regions (CDRs) against Reg4 were assigned using MOE and ZDOCK servers. The binding affinity (KD) was determined by bio-layer interferometry (BLI). MKN45 and AGS cell proliferation was determined by Thiazolyl blue tetrazolium bromide (MTT) method and the cell apoptosis was detected by flow cytometry assay. RESULTS: The KD of scFv-Reg4 to Reg4 was determined to be 1.91×10-8. In MKN45 and AGS cell lines, scFv- Reg4 depressed Reg4-stimulated cell proliferation and the inhibitory rates were 27.7±1.5% and 17.3±2.6%, respectively. Furthermore, scFv significantly enhanced 5-FU-induced cell death, from 23.0±1.0% to 28.4±1.2% in MKN45 and 28.2±0.7% to 36.6±0.6% in AGS cells. Treatment with scFv alone could lyse cancer cells to a certain extent, but no significance has been observed. CONCLUSION: The single-chain antibody (scFv-Reg4) significantly inhibited gastric cancer cell proliferation and synergistically enhanced the lethal effect of 5-FU. Thus, traditional chemo-/radio- therapeutics supplemented with scFv-Reg4 may provide advances in the strategy for gastrointestinal cancer treatment.