Evaluation of the cross-protection of the Vero cell-derived attenuated influenza vaccines with compound adjuvant, through intranasal immunization.

This study was to evaluate the sufficient safety and effect of the novel influenza vaccine program. It prepared new reassortant influenza virus, with high yield on Vero cells. According to the plaque counting, one dose LAIV was composed with 105 PFU of H1, H3, BY, and BV, respectively. Then mixed this LAIV with compound adjuvant, containing 500 µg/mL of carbopol971P and 50 µg/mL of tetanus toxin. That vaccination was called catt-flu. And it employed the GYZZ02 vaccine (commercialized freeze-dried LAIV, listed in China) as cohort analysis control. All mice received two doses of the vaccine, administered on days 0 and 14, respectively. That catt-flu program could induce more cross-protection with neutralizing antibody against heterogeneous types of influenza virus, not only based on HA but also NA protective antigen, through convenient nasal immunization, which had non-inferiority titter compared with the chicken embryo-derived GYZZ02 vaccine on safe and effect. The Vero cell-derived vaccine (LAIV) combined compound catt adjuvant (contain carbopol971P and tetanus toxin) could provide another safety and protective program of influenza vaccine by intranasal administration, as catt-flu program.

Hypoxic Preconditioning Enhances Cellular Viability and Migratory Ability: Role of DANCR/miR-656-3p/HIF-1α Axis in Placental Mesenchymal Stem Cells.

Preeclampsia (PE) is a common complication of pregnancy characterized by new-onset hypertension, albuminuria, or end-stage organ dysfunction, which is seriously harmful to maternal and infant health. Mesenchymal stem cells (MSCs) are pluripotent stem cells derived from extraembryonic mesoderm. They have the potential for self-renewal, multidirectional differentiation, immunomodulation, and tissue regeneration. Several in vivo and in vitro experiments have confirmed that MSCs can delay the pathological progression of PE and improve maternal and fetal outcomes. However, the major limitations in the application of MSCs are their low-survival rates in ischemic and hypoxic disease areas after transplantation and their low rate of successful migration to the diseased regions. Therefore, enhancing cell viability and migration ability of MSCs in both ischemic and anoxic environments is important. This study aimed to investigate the effects of hypoxic preconditioning on the viability and migration ability of placental mesenchymal stem cells (PMSCs) and their underlying mechanisms. In this study, we found that hypoxic preconditioning enhanced the viability and migration ability of PMSCs, increased the expression of DANCR and hypoxia-inducible factor-1α (HIF-1α), and decreased the expression of miR-656-3p in PMSCs. Inhibiting the expression of HIF-1α and DACNR in PMSCs under hypoxia can inhibit the promotive effect of hypoxic preconditioning on viability and migration ability. In addition, RNA pull down and double luciferase assays confirmed that miR-656-3p could directly bind to DANCR and HIF-1α. In conclusion, our study showed that hypoxia could promote the viability and migration ability of PMSCs through the DANCR/miR-656-3p/HIF-1α axis.

Mediating a host cell signaling pathway linked to overwinter mortality offers a promising therapeutic approach for improving bee health.

INTRODUCTION: Honey bees provides valuable pollination services for world food crops and wild flowering plants which are habitats of many animal species and remove carbon dioxide from the atmosphere, a powerful tool in the fight against climate change. Nevertheless, the honey bee population has been declining and the majority of colony losses occur during the winter. OBJECTIVES: The goal of this study was to understand the mechanisms underlying overwinter colony losses and develop novel therapeutic strategies for improving bee health. METHODS: First, pathogen prevalence in overwintering bees were screened between 2015 and 2018. Second, RNA sequencing (RNA-Seq) for transcriptional profiling of overwintering honey bees was conducted and qRT-PCR was performed to confirm the results of the differential expression of selected genes. Lastly, laboratory bioassays were conducted to measure the effects of cold challenges on bee survivorship and stress responses and to assess the effect of a novel medication for alleviating cold stress in honey bees. RESULTS: We identified that sirtuin signaling pathway is the most significantly enriched pathway among the down-regulated differentially expressed genes (DEGs) in overwintering diseased bees. Moreover, we showed that the expression of SIRT1 gene, a major sirtuin that regulates energy and immune metabolism, was significantly downregulated in bees merely exposed to cold challenges, linking cold stress with altered gene expression of SIRT1. Furthermore, we demonstrated that activation of SIRT1 gene expression by SRT1720, an activator of SIRT1 expression, could improve the physiology and extend the lifespan of cold-stressed bees. CONCLUSION: Our study suggests that increased energy consumption of overwintering bees for maintaining hive temperature reduces the allocation of energy toward immune functions, thus making the overwintering bees more susceptible to disease infections and leading to high winter colony losses. The novel information gained from this study provides a promising avenue for the development of therapeutic strategies for mitigating colony losses, both overwinter and annually.

Polymeric photothermal nanoplatform with the inhibition of aquaporin 3 for anti-metastasis therapy of breast cancer.

Metastasis, as one of major challenges in the cancer treatment, is responsible for the high mortality of breast cancer. It has been reported that breast cancer cell invasion and metastasis are related to aquaporin 3 (AQP3), which is the transmembrane transport channel for H2O2 molecules. Moreover, there is agreement that preventing the metastasis of breast tumor cells in combination with inhibiting the tumor growth is a promising strategy for cancer chemotherapy. Herein, we constructed a flexible photothermal crosslinked polymeric nanovehicle for the delivery of the AQP3 inhibitor, [AuCl2(phen)]+Cl- (Auphen). The polymeric nanovehicle (pOMPC-Dex) is comprised of three modules: 1) pOEGMA-co-pMEO2MA serves as the temperature-responsive segment; 2) pCyanineMA acts as the near-infrared (NIR) optical absorbing motif for photothermal therapy and is conjugated with pOEGMA-co-pMEO2MA to obtain NIR light stimuli-responsive drug release; and 3) pPBAMA-Dex functions as an acidic tumor microenvironment-responsive unit. Auphen was encapsulated into a nanovehicle (Auphen@pOMPC-Dex) through electrostatic interactions. The designed nanoplatform showed a pH- and NIR light stimuli-responsive drug release profile and exhibited the strong inhibition of intracellular H2O2 uptake by breast cancer cells, which led to the inhibition of breast cancer cell migration and invasion in vitro. In a breast cancer mouse model, Auphen@pOMPC-Dex markedly reduced the number of lung metastases in tumor-bearing mice due to the combined suppression of tumor growth and metastasis. Consequently, the fabricated Auphen@pOMPC-Dex may provide a new strategy for the development of comprehensive oncotherapies. STATEMENT OF SIGNIFICANCE: High mortality due to metastasis-induced breast cancer has been a key issue that needs to be addressed. It has been reported that aquaporin 3 (AQP3), a transmembrane transport channel for H2O2 molecules was found to have an accelerated effect on breast cancer cell migration. Hence, a flexible crosslinked polymeric nanoplatform with the inhibition of AQP3 was designed to inhibit metastasis of breast cancer cells. At the same time, we combined suppression of tumor growth with photothermal therapy to enhance the anticancer therapy effect.

An effective detection method for wheat mold based on ultra weak luminescence.

It is widely known that mold is one of important indices in assessing the quality of stored wheat. First, mold will decrease the quality of wheat kernels; the wheat kernels infected by mold can produce secondary metabolites, such as aflatoxins, ochratoxin A, zearalenone, fumonisins and so on. Second, the mycotoxins metabolized by mycetes are extremely harmful to humans; once the food or feed is made of by those wheat kernels infected by mold, it will cause serious health problems on human beings as well as animals. Therefore, the effective and accurate detection of wheat mold is vitally important to evaluate the storage and subsequent processing quality of wheat kernels. However, traditional methods for detecting wheat mold mainly rely on biochemical methods, which always involve complex and long pretreatment processes, and waste part of wheat samples for each detection. In view of this, this paper proposes a type of eco-friendly and nondestructive wheat mold detection method based on ultra weak luminescence. The specific implementation process is as follows: firstly, ultra weak luminescence signals of the healthy and the moldy wheat subsamples are measured by a photon analyzer; secondly, the approximate entropy and multiscale approximate entropy are introduced as the main classification features separately; finally, the detection model has been established based on the support vector machine in order to classify two types of wheat subsamples. The receiver operating characteristic curve of the newly established detection model shows that the highest classification accuracy rate can reach 93.1%, which illustrates that our proposed detection model is feasible and promising for detecting wheat mold.

A Biomimetic Nonantibiotic Nanoplatform for Low-Temperature Photothermal Treatment of Urinary Tract Infections Caused by Uropathogenic Escherichia coli.

Urinary tract infections (UTIs) caused by uropathogenic Escherichia coli (UPEC) remain a matter of concern, as the clinical use of multiple antibiotics induces antibiotic resistance in bacteria, resulting in the failure of treatments. Despite the emergence of anti-adhesion strategies that can prevent the development of bacterial drug resistance, these strategies are mainly used for disease prevention rather than effective treatment. Photothermal therapy (PTT) has emerged as an efficient alternative for the elimination of bacteria. Nevertheless, high local temperatures related to PTT probably cause damage to surrounding healthy tissue. Herein, a biomimetic nonantibiotic nanoplatform for low-temperature photothermal treatment of UTIs is developed. The nanoplatform comprises polydopamine (PDA) photothermal core and biphenyl mannoside (Man) shell with multivalent high-affinity to UPEC. Scanning electron microscope (SEM) shows PDA-Man possessed ultra-strong targeting binding ability toward UPEC. It is the fact that this impulse UPEC to form a large bacterial cluster. Consequently, the high photothermal energy of the PDA-Man appears predominantly in the affected bacterial area, while the overall environment remains at a low temperature. The fabricated nanoplatform shows excellent photothermal bactericidal effects, approximately 100% in a UTI model. Overall, this low-temperature photothermal nanoplatform provides an appropriate strategy for the elimination of bacteria in clinical applications.

A Pooled Study of Angiotensin-Converting Enzyme Insertion/Deletion Gene Polymorphism in Relation to Risk, Pathology and Prognosis of Childhood Immunoglobulin A Vasculitis Nephritis.

The angiotensin-converting enzyme (ACE) insertion/deletion (I/D) gene polymorphism has been inconsistently reported to be a risk factor for Childhood immunoglobulin A vasculitis (IgAV) nephritis. We comprehensively searched electronic databases as of Jan 2020. Nineteen studies with 1104 cases and 1589 controls were included. Sensitivity analyses based on different subgroups were performed. Further analyses were conducted for association of ACE polymorphism with disease severity and prognosis. Significant associations were found between ACE I/D polymorphism and childhood IgAV nephritis, with the strongest association in DD vs. II comparison (OR 1.72, 95% CI 1.21-2.46). Subgroup analyses generally showed significant results. Besides, ACE polymorphism was significantly associated with proteinuria (DD + DI vs. II: OR 2.22, 95% CI 1.14-4.33; DI + II vs. DD: OR 0.49, 95% CI 0.30-0.81) and worse prognosis (the strongest effect in DD + DI vs. II: OR 4.43, 95% CI 1.84-10.71) among children with IgAV nephritis. The ACE polymorphism seemed not to be associated with hematuria, hypertension, and renal pathology. This study suggested significant association of ACE gene polymorphism with the risk of IgAV nephritis in children. D allele in the ACE genotype could be a useful genetic marker to predict proteinuria and worse prognosis for childhood IgAV nephritis.

[Application research of entropy weight-based grey systematic theory in quality evaluation of Angelicae Sinensis Radix slices].

In order to discuss the "entropy weight method" for weighting various indicators in the comprehensive evaluation of Angelicae Sinensis Radix slices(ASR), the quality of ASR was comprehensively evaluated by entropy weight-based gray systematic theory and cluster analysis. In this study, the contents of ferulic acid, volatile oil, polysaccharide, alcohol extract, water extract, moisture, total ash and acid-insoluble ash in 44 batches of ASR from different sources were determined. The entropy weight method was used for objective weighting. With relative correlation(r_i) as a measure, a multi-index comprehensive evaluation model was constructed for the quality of ASR. The results showed that the relative correlation value of 44 batches of ASR ranged from 0.301 9 to 0.662 9. There were certain differences in the quality of ASR from different sources. The ASR S1-S8, traceable and standardized in processing techno-logy, showed a high relative correlation degree and high quality ranking, indicating that the implementation of systemic management of the production chain of Chinese herbal pieces was beneficial to the quality control of ASR. The quality evaluation results of 44 batches of ASR were consistent with those of traditional geo-authentic habitats for ASR and the mainstream varieties of ASR on market, and basically consistent with the results of cluster analysis. This study suggests that the gray systematic theory based on the entropy weighting method can be used for the quality evaluation of ASR. The objective weighting of the entropy weight method improves the reliability of the gray correlation method and the scientificity of ASR quality evaluation.

Effect of cell phone radiation on neutrophil of mice.

Purpose: The present study aims to evaluate the effect of cell phone radiation on neutrophil of mice. Materials and methods: 40 male BALB/C mice were randomly divided into four groups as control, blank control, TD-CDMA, and LTE-advanced groups, respectively. Mice were exposed to cell phone radiation for a period of 6 weeks. Then numbers of neutrophil were detected by fully automatic hematology analyzer. Soft agar diffusion method was performed to assess the chemotaxis of neutrophils while the phagocytosis of neutrophils was determined by measuring the staphylococcus albus phagocytosis percentage. Apoptosis was analyzed by flow cytometry. Results: No significant differences were observed among the control and exposure groups regarding the numbers of neutrophils after 2 weeks' exposure to cell phone radiation, while the numbers of neutrophils in TD-SCDMA and LTE-advanced groups were seen to rise after an exposure of 4 or 6 weeks. No effect was observed on chemotaxis of neutrophils due to phone radiation. The phagocytosis of neutrophils was decreased while the apoptosis were increased both in TD-SCDMA and LTE-advanced groups after 6 weeks exposure. Conclusions: Mobile phone radiation could give rise to increase of neutrophil numbers yet with no effect whatever on neutrophils chemotaxis, and the radiation was likely to cause decrease of phagocytosis and induced apoptosis of neutrophils.

Broad-band and high-efficiency polarization converters around 1550 nm based on composite structures.

Broad-band and high-efficiency polarization converter is an imperative component in communication systems, but its functionality often clashes with the constraint of materials. Herein we theoretically and numerically demonstrate that a broad-band and high-efficiency 90° polarization rotator around 1550 nm can be realized using an ultrathin and geometry-optimized composite structure. Based on simulation results, the reflection efficiency and operation bandwidth is up to ≈80% and ≈300 nm, respectively, for the 90° polarization rotator. With similar concept, we also demonstrate a quarter-wave plate with an efficiency of 94% and bandwidth of 110 nm. The electric filed distribution indicates that the conversion behaviors are caused by the strong magnetic coupling in the designed composite structure. Furthermore, the polarization ellipticity properties are investigated to further understand the broad-band effect of the proposed polarization convertors.

Comparative Pharmacokinetic Profiles of Three Protoberberine-type Alkaloids from Raw and Bile-processed Rhizoma coptidis in Heat Syndrome Rats.

BACKGROUND: The Bile-processed Rhizoma coptidis (BRC), which has a colder drug property than Rhizoma coptidis (RC), is widely used for the treatment of heat syndrome. We compared the pharmacokinetics of the protoberberine-type alkaloids in BRC and RC in rats with heat syndrome to elucidate the bile-processing mechanism. MATERIAL AND METHODS: We established a rapid and sensitive method for simultaneously determining three alkaloids: berberine, palmatine, and jatrorrhizine, in rat plasma based on ultra-performance liquid chromatography/tandem mass spectrometry. The separation was carried out on a Waters ACQUITY BEA C18 column. The mobile phase consisted of acetonitrile (containing 0.1% formic acid) and water (containing 0.1% formic acid and 10 mmol/L ammonium acetate) and carbamazepine was used as an internal standard. The detection was carried out in a multiple reaction monitoring mode (MRM) using electrospray ionization in the positive ion mode. RESULTS: Pharmacokinetic profiles indicated that the Cmax of berberine and palmatine increased two times and the Tmax of the three alkaloids decreased three times after bile processing. AUC0→∞ and AUC0→t of the alkaloids were similar between RC and BRC. CONCLUSION: The results suggest that bile processing could increase the absorption rate of alkaloids. This study broadens our understanding of Chinese herbal medicine processing. SUMMARY: Contents of berberine, palmatine and jatrorrhizine, in heat syndrome rats' plasma between the raw and bile-processed Rhizoma coptidis (RC) were determined by UPLC-MS/MS.The whole pharmacokinetic profiles of three alkaloids in the bile-processed Rhizoma coptidis (BRC) were similar to those of RC.The shorter Tmax and increased 2-fold Cmax were obtained after RC bile-processing.Bile-processing could promote the absorption rate of alkaloids in a certain degree. Abbreviation Used: RC: Rhizoma coptidis, BRC: Bile-processed Rhizoma coptidis, HPLC: high-performance liquid chromatography, UPLC-MS/MS: ultra-performance liquid chromatography-mass spectrometry/ mass spectrometry, LC-MS: liquid chromatography-mass spectrometry, MRM: multiple reaction monitoring mode, QC: quality control, RE: relative error, RSD: relative standard deviation, Cmax: maxium of drug concentration, Tmax: time for maxium of drug concentration, AUC: area under concentration-time curve, LLOQ: Linearity and lower limits of quantification, t1/2: half-life, Clz: body clearance.

The memory enhancement effect of Kai Xin San on cognitive deficit induced by simulated weightlessness in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: It is vital for astronauts to develop effective countermeasures to prevent their decline of cognitive performance in microgravity to make space-flight missions successful. The traditional Chinese herbal formula Kai Xin San (KXS) has been used to treat amnesia for thousands years. It is a traditional complex prescription comprising of ginseng (Panax ginseng C. A. Meyer), hoelen (Poria cocos (Schw.) Wolf), polygala (Polygala tenaifolia Willd), and acorus (Acorus tatarinowii Schott). Previous study showed KXS could improve CMS-induced memory impairment in rats. MATERIAL AND METHODS: In this paper, a unique environmental factor-microgravity (weightlessness) was simulated as hindlimb suspension (HLS) by tail in rats for two weeks as the HLS animal model. The KXS at the doses of 0.3 or 0.6g/kg p.o. daily was administrated to HLS rats for two weeks at the same time of HLS, the memory behavior tests were investigated with Morris water maze (MWM) and Shuttle Box (SB) test. The levels of ROS, 8-OHdG and 3-nitrotyrosine (3-NT) in the serum, and AChE and ChAT activity in the brain of rats were determined by ELISA or biochemical analysis. RESULTS: After HLS for two weeks, the escape latency and the swimming distance were significantly increased in the MWM test in rats in the HLS group, compared with control group. The percent of swimming distance in target quadrant and the number of target crossing was significantly decreased in rats in the HLS group compared with the control group. Performance in the SB test showed, the numbers and the distance of active avoidance was decreased from day 4 to day 7, the time spent in electric area was increased in rats in the HLS group compared with the control group. Administration of KXS 0.3 or 0.6g/kg to the HLS rats for two weeks significantly reduced the escape latency and the swimming distance, increased the percentage of swimming distance in target quadrant and the number of target crossings (P<0.01, compared with the HLS group) in the MWM test. Similar treatment with KXS increased the numbers and the distance of active avoidance (P<0.01, compared with the HLS group) and reduced the time spent in electric area after training 3 days in the SB test (P<0.01, compared with the HLS group). The HLS induced the increase of the ROS, 8-OHdG and 3-NT in the serum of rats, but has little influence on the AChE, ChAT activity in the brain. Only the AChE activity in the cortex and the ChAT activity in the hippocampus had some changes in rats in the HLS model group. After administration of KXS 0.6g/kg for two weeks, the abnormal levels of ROS, 8-OHdG, 3-NT were found reversed in the serum of rats (P<0.05, compared with HLS model group). And KXS 0.3g/kg was found reversed the increased AChE activity in the cortex. CONCLUSIONS: Experimental results from this study show that KXS may improve memory deficiency induced by HLS, its mechanisms are major related to antioxidant activities, rather than the central cholinergic system.

Molecular dynamics simulation to rationalize regioselective hydroxylation of aromatic substrates by soluble methane monooxygenase.

Soluble methane monooxygenase (sMMO) is a bacterial multicomponent enzyme that oxidizes a diverse range of substrates, including aromatic hydrocarbons. We have investigated enzyme-substrate interactions that govern oxidation regioselectivity at various sites of aromatic compounds using substrate docking and molecular dynamics (MD) simulations. Here, we studied the hydroxylation of toluene and ethyl benzene by two forms of Methylosinus trichosporium OB3b (sMMO), that is, wild-type (WT) and two active site mutants (L110Y/G). The two substrates, toluene and ethyl benzene, were docked into the active site of the WT and the L110Y/G mutant models of M. trichosporium OB3b sMMO using the available X-ray structure (PDB id 1 MHZ). The trends observed in the formation of the experimental product were highly correlated with the results obtained from the relatively short MD simulation. These results show that our approach could be an attractive computational tool to rationalize the prediction of product ratios and specificities.

Rapid diagnosis and discrimination of bacterial meningitis in children using gram probe real-time polymerase chain reaction.

In this study, we developed a method of simultaneous detection and discrimination of bacteria in cerebrospinal fluid (CSF) with gram probe real-time polymerase chain reaction (PCR). Our results showed 25 clinical strains representing 13 gram-positive and 12 gram-negative bacterial species. They were identified correctly with the corresponding gram probe. The standard curve showed that the amplification efficiency of templates with different concentrations of bacteria was almost the same with a potential detection limit of 10 colony-forming units/mL. A total of 482 children who were clinically suspected of bacterial meningitis were included in this study. A total of 1.0 mL of CSF was collected from every child and was subjected to gram probe-based PCR (GP-PCR), CSF culture, and CSF routine analysis. The positive rate of the GP-PCR array was (32/482, 6.64%) significantly higher than that of CSF culture (23/482, 4.77%). GP-PCR was proved to be an excellent technique for rapid and accurate diagnosis and discrimination of bacterial meningitis, and hence its use as a diagnostic tool in future seems very promising.

Recombinant HCMV UL128 expression and functional identification of PBMC-attracting activity in vitro.

Human cytomegalovirus (HCMV) has evolved several immune evasion strategies. One strategy is controlling the movement of peripheral blood mononuclear cells (PBMCs) by encoding homologues of chemokines. Our aim was to determine whether HCMV open reading frame (ORF) UL128 could encode a protein that attracts PBMCs like a ß-chemokine. The recombinant UL128 protein was synthesized by construction of a stably transfected CHO-UL128 cell line, and a chemotaxis assay showed that UL128 was able to attract PBMCs with a potency equal to that of MIP-1α in vitro. We hypothesize that UL128 protein may act as a ß-chemokine homologue in viral pathogenesis.

Molecular characterization of tumour heterogeneity and malignant mesothelioma cell differentiation by gene profiling.

Malignant mesothelioma is an aggressive tumour, characterized by a variable differentiation pattern and poor prognosis. At present, the clinical outcome in patients with malignant mesothelioma is mainly predicted by the morphological phenotype of the tumour. However, this conventional clinicopathological parameter is of limited value, partly because of the biological heterogeneity of this tumour and poor understanding of the regulatory mechanisms underlying the various patterns of growth. To elucidate the intrinsic molecular programmes that determine tumour differentiation, oligonucleotide arrays were used in an in vitro model of mesothelioma differentiation. The analysis of 2059 genes detected 102 genes that were significantly deregulated. Clustering of these genes into functional categories showed distinctive patterns for the two phenotypes, namely epithelioid and sarcomatoid. The molecular fingerprint of the sarcomatoid tumour component indicates overrepresentation of growth factor receptors and growth factor binding proteins, whereas epithelioid mesothelioma cells express other tumour-promoting factors involved in differentiation, metabolism, and regulation of apoptosis. These differences in the molecular phenotype may give a better basis for diagnosis and for designing novel therapies.