Realgar is a naturally occurring arsenic sulfide (or Xionghuang, in Chinese). It contains over 90% tetra-arsenic tetra-sulfide (As4S4). Currently, realgar has been confirmed the antitumor activities, both in vitro and in vivo, of realgar extracted using Acidithiobacillus ferrooxidans (A. ferrooxidans). Bioleaching, a new technology to greatly improve the use rate of arsenic extraction from realgar using bacteria, is a novel methodology that addressed a limitation of the traditional method for realgar preparation. The present systematic review reports on the research progress in realgar bioleaching and its antitumor mechanism as an anticancer agent. A total of 93 research articles that report on the biological activity of extracts from realgar using bacteria and its preparation were presented in this review. The realgar bioleaching solution (RBS) works by inducing apoptosis when it is used to treat tumor cells in vitro and in vivo. When it is used to treat animal model organisms in vivo, such as mice and Caenorhabditis elegans, tumor tissues grew more slowly, with mass necrosis. Meanwhile, the agent also showed obvious inhibition of tumor cell growth. Bioleaching technology greatly improves the utilization of realgar and is a novel methodology to improve the traditional method.

Shengmai (SM) formula, a classical traditional Chinese medicine formula, is composed of Panax ginseng (Pg), Ophiopogon japonicus (Oj), and Schisandra Chinesis (Sc). SM has been clinically used to treat heart failure and ischemic heart disease. Although SM formula has been reported to be potential for fighting against Alzheimer’s disease (AD) by previous works, there are many gaps in our knowledge on its usage in AD treatment on an organism level and will then need to be further clarified. In this study, transgenic Caenorhabditis elegans expressing human Aβ1–42 are used to evaluate SM formula efficacy to treat AD phenotype and to investigate its underlying mechanism. The results showed that SM formula ameliorated AD pathological characteristics of paralysis behavior and chemotaxis defect in transgenic C. elegans. With SM treatment, the number of Aβ deposits decreased, the levels of gene expressions of hsp16-2, hsp16-41, ace-1, ace-2, and TNFA1P1 homolog genes were down-regulated. Our results also showed that Oj exhibited more stronger effect on delaying paralysis in worms than Pg and Sc did, and synergistic action was observed between Pg and Oj, and Sc further enhanced the activity of Pg/Oj combination on delaying paralysis behavior. Further, SM with herbs of Pg, Oj, and Sc at a dose proportion of 9:9:6 exhibited superior therapeutic efficacy in comparison with herbs at other dose proportions. After SM formula extracted by ethanol, it delayed AD symptoms on a wider dose from 0.2 to 10.0 mg/mL with no toxic effect. These results provided more evidence for SM formula being potential to be used to treat AD.

Neuroinflammation constitutes a principal process involved in the progression of various central nervous system (CNS) disorders, including Parkinson’s disease, Alzheimer’s disease, ischemic stroke, and traumatic brain injury. The safety and efficacy of potential neuroprotective therapeutic agents is controversial and limited. Alpha-melanocyte-stimulating hormone (α-MSH) as a tridecapeptide derived from pro-opiomelanocortin displays potent anti-inflammatory and protective effects with a wide therapeutic window in brain damage. However, it is difficult to deliver effective concentrations of α-MSH into brain tissue via nondirect application. Besides, the half-life of the tridecapeptide is only a few minutes. In the present study, we generated a novel TAT-HSA-α-MSH by genetically fusing α-MSH with N-terminus 11-amino acid protein transduction domain of the human immunodeficiency virus Tat protein (TAT) and human serum albumin (HSA), which showed favorable pharmacokinetic properties and can effectively cross the blood brain barrier (BBB). The findings showed that TAT-HSA-α-MSH significantly inhibits NF-κB activation in human glioma cells A172 and tumor necrosis factor-α (TNF-α) production in experimental brain inflammation. These results indicate that TAT-HSA-α-MSH may be a potential therapeutic agent for treating neuroinflammation which plays a fundamental role in CNS disorders.

The 14-amino acid (IEGPTLRQWLAARA) thrombopoietin mimetic peptide (TMP) shares no sequence homology with native thrombopoietin (TPO). When dimerized, it displays a high-binding affinity for the TPO receptor and has equipotent bioactivity with recombinant human TPO (rhTPO) in stimulating proliferation and maturation of megakaryocytes in vitro. However, TMP is limited for clinical usage because of its short half-life in vivo. In this study, fusion proteins that composed of tandem dimer of TMP (dTMP) genetically fused at the C- or N-terminus of human serum albumin (HSA) were separately expressed in Chinese hamster ovary (CHO) cells. In vitro bioactivity assays showed that purified fusion proteins promoted the proliferation of megakaryocytes in a dose-dependent manner and activated signal transducer and activator of transcription (STAT) pathway in TPO receptor-dependent manner. Following subcutaneous administration, both HSA-dTMP and dTMP-HSA significantly elevated peripheral platelet counts in normal mice in a dose-dependent manner. In addition, fusion with HSA successfully prolonged dTMP half-life in mice. However, when HSA was fused at the C-terminus of dTMP, the bioactivity of dTMP-HSA was about half of that of HSA-dTMP. In conclusion, these results suggested that HSA/dTMP fusion proteins might be potential drugs for thrombocytopenia and, when HSA was fused at the N-terminus of dTMP, the fusion protein had a higher activity.

•Series of novel hydroxycinnamic acid hydrazide were synthesized.•The anticancer activity was evaluated by MTT methods.•Compound 2c induced cell cycle arrest at G2/M phase in H1299 cells.•Caspase-3 mediated pathway is involved in 2c induced apoptosis.In order to generate compounds with superior antitumor activity and reduced toxicity, twelve new hydroxycinnamic acid hydrazide derivatives were synthesized and evaluated for their antiproliferative activities against two cancer cell lines (H1299 lung carcinoma cells and MCF-7 breast cancer cells), and compared to two normal counterparts (NL-20 lung epithelial cells and H184B5F5/M10 breast cells) by MTT method. The results demonstrated that some of these compounds possessed good antiproliferative activity against the two cancer cell lines. Among them, compound 2c was active against the growth of H1299 lung carcinoma cells with IC50 values of 1.50 μM, which was more active than the positive topotecan (IC50 = 4.18 μM). Simultaneously, it showed lower cytotoxic effects on normal NL-20 lung epithelial cells (IC50 > 10 μM). Mechanism studies indicated that it induced cell cycle arrest at G2/M phase followed by activation of caspase-3, and consequently caused the cell death. Further studies on the structure optimization are ongoing.Effects of 2c (1.0–3.0 μM) on the activation of caspase-3. Protein loading was normalized to the expression of β-actin. A representative experiment is shown of three performed.

There is considerable interest in using traditional Chinese medicine formulas (TCMF) to delay aging or treat age-related diseases. Due to cost and duration, the beneficial effects of TCMF on prolongation are mainly extrapolated from vitro studies or physiological indexes. Little is known about whether TCMF are beneficial in whole level, particularly with respect to lifespan. To address this issue, we selected eight formulas with anti-oxidative activity and examined their effects on the lifespan of Caenorhabditis elegans. The results showed that seven of the eight formulas could prolong lifespan of TK22 mutant significantly and five of the eight formulas could obviously extend lifespan of N2 wild-type. To further characterize the prolongation effects, oxidative stress, thermal stress and reproduction test were assayed. We found that the formulas that extended lifespan of TK22 could also protect it from oxidative stress, without reducing the reproductive capacity. Meanwhile, the formulas that prolonged lifespan of N2 wild-type could also enhance its resistance against thermal stress, with damaging the reproductive fitness. These observations indicate that TCMF used in our experiment could be potential therapeutics for anti-aging.

A milk-clotting enzyme named YS-1 was purified from a Bacillus subtilis (B. subtilis) YB-3, which we have isolated from Tibetan Plateau, Gansu, China. The enzyme YS-1 was identified as a metalloproteinase. SDS-PAGE and MALDI-TOF-MS analysis of the purified enzyme gave a molecular weight of 42 kDa. YS-1 was stable over a wide range of temperature from 20 to 60 °C. Purified YS-1 was also active over a wide range of pH from 5.0 to 9.0. It can be activated by Ca2+ and Al3+ but inhibited by Zn2+, Fe2+ and Cu2+. The milk-clotting enzyme YS-1 exhibited high specificity to substrate β-casein and yak milk casein and led to a 75% more rapid coagulation of yak milk than cow milk, due to high β-casein content in yak milk. Together, our findings confirmed that the enzyme YS-1 has a potential to be used in yak cheese industry.

To assay hydrogen peroxide (H2O2) production by phytopathogenic bacteria in vivo, cowpea seedlings were inoculated using wild-type Xanthomonas campestris pv. phaseoli and its mutant strain on ahpC (with attenuated H2O2 production). Antibiotic rifampicin (Rif) was used to kill off the Xanthomonas campestris pv. phaseoli in cowpea leaves. The optimal concentration of Rif to eradicate the bacteria was 500 μg mL–1, and duration of the Rif treatment to eradicate the bacteria in the cowpea tissues was 48 h. This level of Rif did not significantly affect chloroplast and mitochondrial function in cowpea leaves. The concentration of H2O2 in cowpea inoculated with wild-type or mutant strain of Xanthomonas campestris pv. phaseoli following Rif treatment was measured. Increased H2O2 accumulation was observed in cowpea leaves respectively inoculated with wild-type and mutant strain. The accumulation of excess H2O2 was significantly decreased by the Rif treatment on cowpea wild-type bacteria systems. However, the level of H2O2 in cowpea leaves inoculated with mutant strains following Rif treatment was at the same level as in plant leaves without Rif treatment. The results demonstrate that the change in H2O2 concentration observed in vivo was caused by wild-type bacteria.

Magnetite nanocrystal has been extensively used in biomedical field. Currently, an interesting alternative to synthetic magnetic Fe3O4 nanoparticles, called magnetosome, has been found in magnetotactic bacteria. It has been reported that Acidithiobacillus ferrooxidans (At. ferrooxidans) has a potential to synthesize magnetosome. In this study, transmission electron microscope (TEM) was used to analyze the magnetite particles in At. ferrooxidans BY-3. The magnetosomes formed by this bacterium were isolated by a method combining ultracentrifugation and magnetic separation. Crystalline phase and surface functional group of the magnetosomes were investigated by X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR), respectively. Biocompatibility of the magnetosomes was systematically evaluated at various concentrations (0.5, 1.0, 2.0 and 4.0 mg/ml). MTT test, hemolysis assay and Micronucleus Test were carried out to evaluate in vitro cytotoxicity, blood toxicity and genotoxicity of magnetosomes, respectively. Under these conditions, magnetosomes showed no cytotoxic, genotoxic and hemolytic effects up to 4.0 mg/ml indicating good biocompatibility of these biological nanoparticles. These revealed that the magnetosomes might have a potential for biotechnological and biomedical applications in the future.Highlights► The production of magnetosomes from At. ferrooxidans has been easily available. ► Several techniques are used to characterize properties of the magnetosomes. ► The magnetosomes have no cytotoxicity, no hemolysis activity and no genotoxicity.

Ritter reaction of podophyllotoxins with chloroacetonitrile and subsequent cleavage of the chloroacetyl group in the resulting chloroacetamide with thiourea under both classical heating and ultrasonic conditions is an efficient procedure for the synthesis of 4β-aminopodophyllotoxins. In general, significant improvements in the rates of reaction and yields of the sonochemical reactions relative to the classical heating reactions were observed.

We investigated the differential expression of AOX1 multi-gene family and the regulation of alternative respiratory pathway during initial greening development in leaves of rice (Oryza sativa L.) seedlings. After exposing the dark-grown rice seedlings to continuous irradiation, total respiration (Vt), capacity of alternative pathway (Valt), and their ratio (Valt/Vt) increased with the greening of leaves. In this process, AOX1c transcript increased under constant irradiation, while AOX1a and AOX1b transcripts were hardly detected. Thus AOX1c in rice presents a similar expression pattern as AOX2 does in many dicotyledonous species during greening development. Compared with the rapid increase of cyanideresistant respiration in the presence of photon energy, CO2 fixation was not observed until 8 h after the onset of irradiation. The AOX inhibitor salicylhydroxamic acid (SHAM; 1 mM) inhibited 67.3 % of cyanide-insensitive oxygen uptake in dark-grown leaves and 69.4 % of it in leaves grown under irradiation. Dark-grown plants pre-treated with SHAM were then irradiated for 12 h. SHAM did not obviously modify photosynthetic CO2 fixation rate on a chlorophyll (Chl) content basis in both leaves and simultaneously isolated chloroplasts. Hence during initial greening steps of the plants, the induction of alternative pathway and AOX1 expression by irradiation is not directly linked with carbon assimilation of photosynthesis. The application of SHAM partially limited Chl production in rapidly greening leaves, indicating that Chl synthesis in the process of greening might be medicated to some extent by alternative respiratory pathway.

A new Acidithiobacillus ferrooxidans cell immobilization technique utilizing the complex of PVA solution and sodium alginate solution crosslinked by Ca(NO3)2 as entrapment medium is reported. The mixture of A. ferrooxidans suspension and the entrapment complex were extruded into a solution of Ca(NO3)2 (1–5%) to form beads, then the beads were frozen at −20 °C for 1–2 days and thawed at room temperature. The forming mechanism, characteristic of this immobilized beads and the factors affecting activity of immobilized cells were also discussed. A maximum oxidation rate of 4.6 g Fe2+/(L h) was achieved in batch cultures by these immobilized cells. Precipitation formed during culture process was analyzed. The forming mechanism of this precipitation and how this precipitation affects the whole system were also discussed. In addition, the immobilization technique is operated simply, and the gel beads have high stability even under non-sterile conditions. So its application on an industrial scale would be more practicable.

In an attempt to find the biorational pesticides, a series of novel camptothecin derivatives were synthesized via a simple modification of the carbodiimide method using the combination of scandium triflate (Sc(OTf)3) and 4-dimethylaminopyridine (DMAP), and their structures were identified by IR, ESR, HRMS and 1H NMR. Their antifeedant effect and insecticidal activity against third-instar larva of Mythimna separate were examined. The results of preliminary bioassays showed that these compounds exhibited less potent antifeedant activity than camptothecin. Also, all of these derivatives of camptothecin showed delayed insecticidal activity, which differs from traditional neurotoxic insecticides. The results suggested that 20-hydroxyl group in the camptothecin derivatives is essential to keep the insecticidal activity and nature of the substitutes has a major impact on insecticidal activity of these compounds.

The contents of free salicylic acid and conjugated salicylic acid were determined by using TLC and HPLC at different time after attack by Tetranychus urticae Koch in the first true leaf of frijole (Phaseolus vulgaris. L) plant. A maximal nine fold increase in the free salicylic acid content in the attacked leaves was observed. The increase of conjugated salicylic acid was also significant. The content of free salicylic acid in the systemic leaves increased about 500%, while the content of conjugated salicylic acid in the systemic leaves increased by two to three times.

Serofluid dish (or Jiangshui, in Chinese), a traditional food in the Chinese culture, is made from vegetables by fermentation. In this study, bacterial community of the fermented serofluid dish was assessed by Illumina amplicon sequencing. The metagenome comprised of 49,589 average raw reads with an average 11,497,917 bp and G + C content is 52.46%. This is the first report on V4 hyper-variable region of the 16S rRNA metagenome sequence employing Illumina platform to profile the microbial community of this little known fermented food from Gansu Province, China. The metagenome sequence can be accessed at NCBI, SRA database accession no. SRP065370.

Serofluid dish (or Jiangshui, in Chinese), a traditional food in the Chinese culture for thousands of years, is made from vegetables by fermentation. In this work, microorganism community of the fermented serofluid dish was investigated by the culture-independent method. The metagenomic data in this article contains the sequences of fungal internal transcribed spacer (ITS) regions of rRNA genes from 12 different serofluid dish samples. The metagenome comprised of 50,865 average raw reads with an average of 8,958,220 bp and G + C content is 45.62%. This is the first report on metagenomic data of fungal ITS from serofluid dish employing Illumina platform to profile the fungal communities of this little known fermented food from Gansu Province, China. The Metagenomic data of fungal internal transcribed spacer can be accessed at NCBI, SRA database accession no. SRP067411.

By four different methods, Acidithiobacillus ferrooxidans cells were immobilized by the complex of PVA and sodium alginate. The beads formed by these different methods were evaluated in terms of relative mechanical strength, biological activity, dilatability, and so on. The results indicate that the technique utilizing the complex of PVA and sodium alginate crosslinked with Ca(NO3)2 is more appropriate for the immobilization of A. ferrooxidans than any others. So the PVA–calcium nitrate beads were used in batch and continuous culture. A maximum ferrous iron oxidation rate of 4.6 g/l/h was achieved in batch culture. Long-time performance of packed-bed bioreactor was evaluated systematically over 40 days, depending on the conversion ratio of ferrous iron and the residence time. At a residence time of 2.5 h, 96% of the initial ferrous iron was oxidized. This study shows this new immobilization technique will be a feasible and economical method for A. ferrooxidans.

Acidithiobacillus ferrooxidans type strain DLC-5, isolated from Wudalianchi in Heihe of Heilongjiang Province, China. Here, we present the draft genome of strain DLC-5 which contains 4,232,149 bp in 2745 contigs with 57.628% GC content and includes 32,719 protein-coding genes and 64 tRNA-encoding genes. The genome sequence can be accessed at DDBJ/EMBL/GenBank under the accession no. JNNH00000000.1.

Hepatic oxidative stress plays a key role in the development of non-alcoholic steatohepatitis (NASH), therefore, treatment approaches that address the antioxidant is helpful in the therapy of patients with NASH. N-trans-coumaroyloctopamine (1) and N-trans-feruloyloctopamine (2) were identified as the primary antioxidant constituents of garlic skin with high antioxidant activities. The aim of this study was to elucidate the protective effect and mechanism of the antioxidants on NASH in rats. The results provide morphological and molecular biological evidences for the protective role of the antioxidant 2 in ameliorating oxidative stress and hepatic apoptosis in experimental NASH for the first time. Mechanism study indicated that the antioxidant 2 significantly reduced the expression of COX-2 mRNA and protein by western blot, RT-PCR and immunohistochemical techniques.

•The problem of un-dissolvable of realgar is resolved by bio-transformation.•Bio-transformation was influenced by extracellular substance.•The extracellular substances affect the content of arsenic bio-transformation.•The study reveals the novel application of Acidithiobacillus ferrooxidans.Due to the toxic and un-dissolvable properties, the medical application of realgar (As4S4) has always been limited for a long time. In this study, a novel method of dissolving realgar by immobilized Acidithiobacillus ferrooxidans (A. ferrooxidans) is introduced. The characterizations of dissolving realgar are carried out by means of SEM, ICP-AES, XRD and FTIR. The results suggest that the bio-transformation process is influenced by extracellular substances such as pH value and the oxidation of Fe2 +. These extracellular substances affect the concentration of arsenic in bio-transformation solution rather than A. ferrooxidans. Present study reveals the novel application of A. ferrooxidans.