Development of a Safe and Effective Bacillus thuringiensis-Based Nanobiopesticide for Controlling Tea Pests.

Mg(OH)2 was used as the nanocarrier of the Bacillus thuringiensis (Bt) Cry1Ac protein, and the synthesized Cry1Ac-Mg(OH)2 composites were regular and uniform nanosheets. Nano-Mg(OH)2 could effectively improve the insecticidal effect of the Cry1Ac protein toward Ectropis obliqua. It could enhance the damage degree of the Cry1Ac protein to intestinal epithelial cells and microvilli, induce and enrich the production of reactive oxygen species (ROS) in the midgut, and enhance the degradation of the Cry1Ac protein into active fragments. Furthermore, an anti-rinsing assay showed that the Cry1Ac-Mg(OH)2 composites were bound to the notch structure of the tea leaf surface. The retention of the Cry1Ac protein increased by 11.45%, and sprayed nano-Mg(OH)2 was rapidly absorbed by different tissues of tea plants. Moreover, nano-Mg(OH)2 and composites did not significantly affect non-target organisms. These results show that nano-Mg(OH)2 can serve as a safe and effective biopesticide carrier, which provides a new approach for stable and efficient Bt preparation.

Mesoporous Silica Nanoparticles Induce Intracellular Peroxidation Damage of Phytophthora infestans: A New Type of Green Fungicide for Late Blight Control.

Nanopesticides are considered to be a promising alternative strategy for enhancing bioactivity and delaying the development of pathogen resistance to pesticides. Here, a new type of nanosilica fungicide was proposed and demonstrated to control late blight by inducing intracellular peroxidation damage to Phytophthora infestans, the pathogen associated with potato late blight. Results indicated that the structural features of different silica nanoparticles were largely responsible for their antimicrobial activities. Mesoporous silica nanoparticles (MSNs) exhibited the highest antimicrobial activity with a 98.02% inhibition rate of P. infestans, causing oxidative stress responses and cell structure damage in P. infestans. For the first time, MSNs were found to selectively induce spontaneous excess production of intracellular reactive oxygen species in pathogenic cells, including hydroxyl radicals (•OH), superoxide radicals (•O2-), and singlet oxygen (1O2), leading to peroxidation damage in P. infestans. The effectiveness of MSNs was further tested in the pot experiments as well as leaf and tuber infection, and successful control of potato late blight was achieved with high plant compatibility and safety. This work provides new insights into the antimicrobial mechanism of nanosilica and highlights the use of nanoparticles for controlling late blight with green and highly efficient nanofungicides.

Oxygen therapy accelerates apoptosis induced by selenium compounds via regulating Nrf2/MAPK signaling pathway in hepatocellular carcinoma.

Selenium has good antitumor effects in vitro, but the hypoxic microenvironment in solid tumors makes its clinical efficacy unsatisfactory. We hypothesized that the combination with oxygen therapy might improve the treatment efficacy of selenium in hypoxic tumors through the changes of redox environment. In this work, two selenium compounds, Na2SeO3 and CysSeSeCys, were selected to interrogate their therapeutic effects on hepatocellular carcinoma (HCC) under different oxygen levels. In tumor-bearing mice, both selenium compounds significantly inhibited the tumor growth, and combined with oxygen therapy further reduced the tumor volume about 50 %. In vitro HepG2 cell experiments, selenium induced autophagy and delayed apoptosis under hypoxia (1 % O2), while inhibited autophagy and accelerated apoptosis under hyperoxia (60 % O2). We found that, in contrast to hypoxia, the hyperoxic environment facilitated the H2Se, produced by the selenium metabolism in cells, to be rapidly oxidized to generate H2O2, leading to inhibit the expression level of Nrf2 and to increase that of phosphorylation of p38 and MKK4, resulting in inhibiting autophagy and accelerating apoptosis. Once the Nrf2 gene was knocked down, selenium compounds combined with hyperoxia treatment would further activate the MAPK signaling pathway and further increase apoptosis. These findings highlight oxygen can significantly enhance the anti-HCC effect of selenium compounds through regulating the Nrf2 and MAPK signaling pathways, thus providing novel therapeutic strategy for the hypoxic tumors and pave the way for the application of selenium in clinical treatment.

Correlation between vaginal flora and cervical immune function of human papilloma virus-infected patients with cervical cancer.

Background: To analyse the correlation between vaginal flora and cervical immune function of HPV-infected patients with cervical cancer. Methods: Six hundred females with genital tract infections treated in Xuzhou Hospital of Traditional Chinese Medicine from January 2014 to December 2016 were selected and divided into a high-risk HPV group (n=246) and a control group (n=354). The vaginal flora and human T lymphocyte subsets (CD3+, CD4+, CD8+) were detected. Multivariate logistic regression analysis was performed to explore the risk factors for HPV infection. Results: The numbers of CD4+ and CD4+/CD8+ T cells of the high-risk HPV group were significantly lower than those of the control group (P<0.05). The two groups had similar numbers of CD3+ and CD8+ T cells. In the high-risk HPV group, the positive rates of Lactobacillus, Chlamydia trachomatis, Mycoplasma hominis, mycetes, Ureaplasma urealyticum and bacterial vaginosis were significantly higher than those of the control group (P<0.05). There was no significant difference in the positive rates of trichomonads between the two groups. Multivariate logistic regression analysis revealed that C. trachomatis and U. urealyticum were independent risk factors for high-risk HPV infection (P<0.05). Conclusion: High-risk HPV infection in patients with cervical cancer was associated with vaginal flora and immune function. C. trachomatis and U. urealyticum were independent risk factors for high-risk HPV infection.

Single-cell infrared phenomics: phenotypic screening with infrared microspectroscopy.

We conceptually demonstrate single-cell infrared phenomics as a novel strategy of phenotypic screening with infrared microspectroscopy. Based on this development, the cancer cell HepG2 glycocalyx was first identified as a potential target of protopanaxadiol, an herbal medicine. These findings provide a powerful tool to accurately evaluate the cell stress response and to largely expand the phenotypic screening toolkit for drug discovery.

Temperature-sensitive hydrogel for rectal perfusion improved the therapeutic effect of Kangfuxin liquid on DSS-induced ulcerative colitis mice: The inflammation alleviation and the colonic mucosal barriers repair.

Kangfuxin liquid (KFX) is a Chinese medicine extracted from Periplaneta americana dried worms, which presented the bioactive functions of anti-inflammation and promoting the gastrointestinal mucosal barriers repair. But the low availability of KFX exposed to the distal colon affects its therapeutic effect on ulcerative colitis. Herein, an in situ hydrogel containing KFX was designed by using temperature-sensitive poloxamer 407 (P-407) as material for rectal administration. Three KFX-P formulations with different P407 concentrations (17%, 20% and 25%) were designed and screened by detecting the gelation time, gelation temperature and mechanical strength of hydrogel. P407 in these formulations was able to be completely dissolved in KFX at 4 ℃ and then was in situ gelled at 37 ℃ to form a semisolid hydrogel. Moreover, the gelation time, the gelation temperature and the mechanical strength of KFX-P hydrogel are highly dependent on P407 concentration. With P407 concentration increasing, both the gelation time and gelation temperature of KFX-P accordingly decreased and the gelation temperature range becomes narrowed; while the mechanical strength increased. KFX-P-20% displayed the moderate gelation temperature (28-30 ℃), the short gelation time (26 s) and the moderate mechanical strength (G' = 4.2 × 103 Pa), which was chosen for animal study. Thereafter, ulcerative colitis mice model (UC) was established by dextran sulfate sodium (DSS) and the therapeutic effect of KFX-P on UC was evaluated by inflammation symptoms relief, colon length, colonic MPO level and colonography. After rectal administration of KFX or KFX-P, the symptoms including diarrhea and hematochezia (DAI scores), weight loss and spleen swelling were significantly hindered. Meanwhile, the colonic MPO level in these groups was significantly decreased in comparison with PBS treatment. But the therapeutic effect of KFX-P was better than KFX. Besides, the morphology and mucosal barrier of colon were evaluated by HE staining, ZO-1 and claudin-5 staining. The mucosa epithelium layer, crypt, muscle layer mucosa and submucosa were also well repaired after KFX-P treatment. The strong fluorescence of ZO-1 and claudin-5 were uniformly distributed along the whole epithelial mucosa after KFX-P treatment, indicating the effective repairing of the colonic mucosal barrier. Collectively, the temperature-sensitive KFX-P for rectal delivery could effectively promote the repair of the colon mucosal barrier and inhibit the colonic inflammation in DSS-induced mice, which may be a potential strategy for UC treatment.

Succinate accumulation induces mitochondrial reactive oxygen species generation and promotes status epilepticus in the kainic acid rat model.

Though succinate accumulation is associated with reactive oxygen species (ROS) production and neuronal injury, which play critical roles in epilepsy, it is unclear whether succinate accumulation contributes to the onset of epilepsy or seizures. We sought to investigate changes in succinate, oxidative stress, and mito-SOX levels, as well as mitophagy and neuronal change, in different status epilepticus (SE) rat models. Our results demonstrate that KA-induced SE was accompanied by increased levels of succinate, oxidative stress, and mito-SOX, as well as mitophagy and neuronal degeneration. The similarly increased levels of succinate, oxidative stress, and mito-SOX were also found in pilocarpine-induced SE. Moreover, the reduction of succinate accumulation by the inhibition of succinate dehydrogenase (SDH), malate/aspartate shuttle (MAS), or purine nucleotide cycle (PNC) served to reduce succinate, oxidative stress, and mito-SOX levels, thereby preventing oxidative stress-related neuronal damage and lessening seizure severity. Interestingly, simulating succinate accumulation with succinic acid dimethyl ester may induce succinate accumulation and increased oxidative stress and mito-SOX levels, as well as behavior and seizures in electroencephalograms similar to those observed in rats exposed to KA. Our results indicate that succinate accumulation may contribute to the increased oxidative stress/mitochondrial ROS levels, neuronal degeneration, and SE induced by KA administration. Furthermore, we found that succinate accumulation was mainly due to the inverse catalysis of SDH from fumarate, which was supplemented by the MAS and PNC pathways. These results reveal new insights into the mechanisms underlying SE and that reducing succinate accumulation may be a clinically useful therapeutic target in SE.

Huganpian, a traditional chinese medicine, inhibits liver cancer growth in vitro and in vivo by inducing autophagy and cell cycle arrest.

Huganpian (HGP), a traditional chinese medicine composed of 6 herbs, possesses excellent therapeutic effects in clinical application. In this study, we aimed to elucidate the anti-tumor activity and the underlying mechanisms of HGP in liver cancer. The results of this study indicated that HGP effectively inhibited liver cancer growth in vitro and in vivo in a dose-dependent manner. Mechanistically, HGP exerted its anti-tumor effects by triggering autophagy with increased LC3Ⅱ and beclin1 levels and arrested the cell cycle on G0-G1 phase by downregulating the expressions of cyclin-dependent kinase 2 (CDK2), cyclin-dependent kinase 4 (CDK4) and cyclinE1 in vitro and in vivo. Meanwhile, HGP did not induce apoptosis significantly. Importantly, we also confirmed that there were fewer side effects of HGP on immune system. Taken together, our findings suggest for the first time that HGP may become a promising drug or adjuvant drug with a lower toxicity for liver cancer treatment in the future.

Shen-Shuai-Ning granule decreased serum concentrations of indoxyl sulphate in uremic patients undergoing peritoneal dialysis.

Clearance of protein-bound uremic toxins (PBUTs) by dialysis is a challenge in the treatment of uremic patients. Shen-Shuai-Ning (SSN), a traditional Chinese medicine formulation, has been used commonly in China to retard kidney disease progression and decrease uremic toxins in chronic kidney disease (CKD) patients, but the effects of SSN on serum PBUTs in dialysis patients were not investigated. We conducted a randomized controlled trial in patients on peritoneal dialysis (PD) at dialysis center of Changzheng Hospital to evaluate the effects of SSN on serum PBUTs. Participants with SSN intervention took 5 g SSN granule three times daily for 12 weeks, while the baseline medications and dialysis prescriptions remained during the study in all patients. The serum concentrations of indoxyl sulphate (IS) and p-cresol sulphate (PCS) were determined by HPLC/MS/MS and biochemical parameters were assessed during the study. Sixty PD patients were enrolled and randomly allocated into SSN group and control group. Total IS level was significantly lower in SSN group than in control group at week 4, 8, and 12 (27.28 ± 18.19, 29.73 ± 19.10, and 29.41 ± 17.61 mg/l compared with 39.25 ± 20.23, 44.86 ± 23.91, and 45.34 ± 33.52 mg/l, respectively). However, there were no statistical difference of total PCS, free forms of IS and PCS concentrations between SSN group and control group during 12 weeks follow-up. Administration of SSN granule orally decreased serum total IS level effectively in uremic patients on PD with good tolerance. Benefits of PD patients' outcomes from IS reduction by SSN awaits further large size and long duration clinical trials to verify.

Flowerlike Mg(OH)2 Cross-Nanosheets for Controlling Cry1Ac Protein Loss: Evaluation of Insecticidal Activity and Biosecurity.

Bacillus thuringiensis (Bt) can produce Cry proteins during the sporulation phase, and Cry protein is effective against lepidopteran, coleopteran, and dipteran insects and nematodes. However, Cry protein tends to be discharged into soil and nontarget plants through rainwater runoff, leading to reduced effective period toward target insects. In the present study, nano-Mg(OH)2 (magnesium hydroxide nanoparticles, MHNPs) were synthesized to control the loss of Cry1Ac protein and deliver protein to Helicoverpa armigera (Lepidoptera: Noctuidae). The results showed that Cry1Ac protein could be loaded onto MHNPs through electrostatic adsorption, and both MHNPs and Cry protein were stable during the adsorption process. Meanwhile, the Cry1Ac-loaded MHNPs could remain on the surface of cotton leaves, resulting in enhanced adhesion of Cry1Ac protein by 59.50% and increased pest mortality by 75.00%. Additionally, MHNPs could be slowly decomposed by acid medium and MHNPs showed no obvious influence on cotton, Bt, Escherichia coli, and H. armigera. Therefore, MHNPs could serve as an efficient nanocarrier for delivery of Cry1Ac protein and be used as a potential adjuvant for biopesticide in agricultural applications.

Effects of Taurine on Alterations of Neurobehavior and Neurodevelopment Key Proteins Expression in Infant Rats by Exposure to Hexabromocyclododecane.

Hexabromocyclododecanes (HBCDs) is a widely used flame retardant. Studies have found that HBCDs has toxic effects on endocrine and neural development, leading to adverse effects on behavior, learning and memory. This study aimed to investigate the protective effects of taurine on cognitive function, neurotrophic factors expression of infant rats exposured to HBCDs. Sprague-Dawley rats of 10-days old were oral gavaged of different doses (0.3, 3 and 30 mg/kg) of HBCDs and 30 mg/kg HBCDs with 300 mg/kg taurine for 60 consecutive days. Rat cognitive function was detected by the method of Morris water maze test. The protein expressions of brain derived neurotrophic factor (BDNF), nerve growth factor (NGF) and fibroblast growth factor (FGF) were assayed by Western-blotting. Results showed that rats exposed to HBCDs significantly declined rats spatial learning and memory ability by increasing the latency time of seeking the platform (P < 0.05), decreasing the numbers that each rat had crossed the non-exits and the time spent in the target quadrant as compared with those in control rats (P < 0.05). Taurine treatment significantly reversed the effects of HBCDs. Western-blotting results showed that expression of BDNF, NGF and FGF proteins in the low dose group were obviously increased compared with those in control rats (P < 0.01), and middle-dose and high dose groups significantly decreased. Taurine treatment increased BDNF and NGF expression as compared with high dose groups while Taurine seemed to have no effects on FGF. These result suggested that higher doses of HBCDs early exposure in the developing rats could decrease neurotrophic factors including BDNF, NGF, FGF, which have an impact on neural development, damage on learning and memory.

Effect of Tai Chi exercise on blood lipid profiles: a meta-analysis of randomized controlled trials.

OBJECTIVE: Studies have demonstrated that Tai Chi exercise improves blood lipid level with inconsistent results. A meta-analysis was conducted to quantify the effects of Tai Chi on blood lipid profiles in humans. METHODS: We screened the databases of PubMed, EMBASE, Cochrane Library (Central), Web of Science, China National Knowledge Infrastructure (CNKI), Wanfang data, and Clinicaltrials.gov for randomized controlled trials with Physiotherapy Evidence Database (PEDro) score more than 3 points up to June 2015. Six studies involving 445 subjects were included. Most trials applied 12-week Tai Chi intervention courses. RESULTS: In comparison with the control group, blood triglyceride (TG) level difference between follow-up and baseline was statistically significantly lower in the Tai Chi practicing group (weighted mean difference (WMD) -16.81 mg/dl; 95% confidence intervals (CI) -31.27 to -2.35 mg/dl; P=0.02). A trend to improving total cholesterol (TC) reduction was found with Tai Chi (WMD -7.96 mg/dl; 95% CI -17.30 to 1.39 mg/dl; P=0.10). However, no difference was found in blood low-density lipoprotein cholesterol (LDL-C) or high-density lipoprotein cholesterol (HDL-C). CONCLUSIONS: Tai Chi exercise lowered blood TG level with a trend to decrease blood TC level. Our data suggest that Tai Chi has the potential to implement meaningful blood lipid modification and serve as an adjunctive exercise modality. The relationship between Tai Chi exercise regimen and lipid profile change might have a scientific priority for future investigation.

A nanosensor for in vivo selenol imaging based on the formation of Au-Se bonds.

Selenol is a key metabolite of Na2SeO3 and plays an important role in many physiological and pathological processes. The real-time monitoring of selenol is of scientific interest for understanding the anti-cancer mechanism of Na2SeO3. Based on selenol's ability to specifically break AuS bonds and form more stable AuSe bonds on the surfaces of gold nanoparticles (AuNPs), we developed a novel near-infrared fluorescent nanosensor (Cy5.5-peptide-AuNPs) for detecting selenol. The nanosensor exhibited rapid response to selenol with high selectivity and sensitivity, and it was successfully used to image changes in the selenol level in HepG2 cells during Na2SeO3-induced apoptosis. Moreover, in vivo fluorescence imaging of selenol was obtained from H22 tumor-bearing mice injected with both the nanosensor and sodium selenite. The results showed that the tumor cell apoptosis induced by Na2SeO3 is correlated with high-level of selenol under hypoxic conditions. We believe that this nanosensor could serve as a powerful tool for monitoring selenol and exploring the physiological function of selenol in a variety of physiological and pathological contexts and that the probe-designed strategy will provide a new platform for research on relevant selenium chemistry.

The mechanism of uranium transformation from U(VI) into nano-uramphite by two indigenous Bacillus thuringiensis strains.

The mechanism of uranium transformation from U(VI) into nano-uramphite by two indigenous Bacillus thuringiensis strains was investigated in the present work. Our data showed that the bacteria isolated from uranium mine possessed highly accumulation ability to U(VI), and the maximum accumulation capacity was around 400 mg U/g biomass (dry weight). X-ray powder diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FT-IR) analyzes indicated that the U(VI) was adsorbed on the bacterial surface firstly through coordinating with phosphate, CH2 and amide groups, and then needle-like amorphous uranium compounds were formed. With the extension of time, the extracellular crystalline substances were disappeared, but some particles were appeared in the intracellular region, and these particles were characterized as tetragonal-uramphite. Moreover, the disrupted experiment indicated that the cell-free extracts had better uranium-immobilization ability than cell debris. Our findings provided the understanding of the uranium transformation process from amorphous uranium to crystalline uramphite, which would be useful in the regulation of uranium immobilization process.

Acupuncture modulates spontaneous activities in the anticorrelated resting brain networks.

Neuroimaging studies of acupuncture have demonstrated extensive signal attenuations in the core regions of a "default mode" network as well as signal potentiations in the regions of a "central-executive" network. We proposed that this acupuncture-related dichotomy may represent the anticorrelation in these intrinsic brain networks showing spontaneous fluctuations during rest. According to a plentiful clinical report, acupuncture can provide pain relief beyond the time it is being administrated; therefore, imaging its sustained effect (rather than acute effect) on the brain networks may further help elucidate the mechanisms by which acupuncture achieves its therapeutic effects. As an interface, the anterior insula (AI) has recently been shown to be a network hub, which initiates dynamic switching between these intrinsic networks. Here, we attempt to explore how acupuncture can modulate spontaneous coherences of these resting networks anchored by the AI. Using a spontaneous activity detection approach, we identified an AI-related dichotomy showing spontaneous activations in the CEN along with wide spontaneous deactivations located exclusively in the DMN. Following verum acupuncture, but not sham control, there was a prominently enhanced dichotomy in the CEN and DMN networks. More importantly, a long-lasting effect of acupuncture could further modulate intrinsic coherences of the wide interoceptive-autonomic areas, including the paralimbic regions and brainstem nuclei. These findings suggested that acupuncture may not only enhance the dichotomy of the anticorrelated resting networks, but also modulate a larger spatio-temporal extent of spontaneous activities in the salient interoceptive-autonomic network, contributing to potential actions in the endogenous pain-modulation circuits and homeostatic control mechanisms.

FMRI connectivity analysis of acupuncture effects on an amygdala-associated brain network.

BACKGROUND: Recently, increasing evidence has indicated that the primary acupuncture effects are mediated by the central nervous system. However, specific brain networks underpinning these effects remain unclear. RESULTS: In the present study using fMRI, we employed a within-condition interregional covariance analysis method to investigate functional connectivity of brain networks involved in acupuncture. The fMRI experiment was performed before, during and after acupuncture manipulations on healthy volunteers at an acupuncture point, which was previously implicated in a neural pathway for pain modulation. We first identified significant fMRI signal changes during acupuncture stimulation in the left amygdala, which was subsequently selected as a functional reference for connectivity analyses. Our results have demonstrated that there is a brain network associated with the amygdala during a resting condition. This network encompasses the brain structures that are implicated in both pain sensation and pain modulation. We also found that such a pain-related network could be modulated by both verum acupuncture and sham acupuncture. Furthermore, compared with a sham acupuncture, the verum acupuncture induced a higher level of correlations among the amygdala-associated network. CONCLUSION: Our findings indicate that acupuncture may change this amygdala-specific brain network into a functional state that underlies pain perception and pain modulation.

Exploratory analysis of functional connectivity network in acupuncture study by a graph theory mode.

Neuroimaging studies involved acupuncture have already demonstrated its significant modulatory effects on the limbic system, paralimbic and subcortical gray structures. These structures with presumably intimated relationships with acupuncture may constitute a network. In this study, we employed a network mode from the graph theory to evaluate the functional connectivity among multiple brain regions in the resting brain. Evidence from our findings clearly supported the existence of a large functional connectivity network constructed during a resting condition with fMRI. We concluded that acupuncture stimulation may induce the modulation of the 'acupuncture-related' network to exert its influence on the therapeutic effects, presented by significant changes of functional connectivity of some brain regions. Our study may provide a further step to the exploration of the underlying mechanism of acupuncture effects.

The correlated network of acupuncture effect: a functional connectivity study.

A functional connectivity, which are temporally correlated in functionally related brain regions, before and after acupuncture manipulation was measured by MRI. Amygdala, as the control system of endogenetic analgesia, was selected for "seed" point. We found that compelling similarity existed in the network of resting state before and after acupuncture manipulation. A paired student t-test was implemented to investigate under the different conditions. The main difference was found in the limbic system, brainstem and cerebellum. We conclude that the default endogenous analgesia functional network exists in human brain at a low level, and it could be increased to a higher level by acupuncture modulation.