Coinoculation of arbuscular mycorrhizal fungi and rhizobia stimulates atrazine dissipation by changing the atrazine-degrading bacterial community at the soil aggregate scale.

As a potential low-cost and environmentally friendly strategy, bioremediation of herbicide polluted soil has attracted increasing attention. However, there is a lack of knowledge regarding the response of the atrazine-degrading bacterial community to coinoculation of arbuscular mycorrhizal (AM) fungi and rhizobia for atrazine dissipation. In this study, a pot experiment was conducted with AM fungi Glomus mosseae (AM), rhizobia Rhizobium trifolii TA-1 (R) and their coinoculation (AMR) with atrazine. In each treatment, the atrazine-degrading bacterial community of four soil size aggregates, namely large macroaggregates (LMa), small macroaggregates (SMa), microaggregates (Mia) and primary particles (P) were investigated. The results showed that the atrazine residue concentration was lowest in AMR, and that in LMa was also significantly lower than that in the other smaller aggregate sizes. Overall, inoculation, the aggregate fraction and their interaction had significant effects on soil TN, SOC, AP and pH. For the atrazine-degrading bacterial community, the Chao1 index increased with decreasing particle size, but the Shannon index decreased. Moreover, the abundances of the dominant atrazine-degrading bacterial genera Arthrobacter, Bacillus, Marmoricola and Nocardioides in the Mia and P particle size groups were greater than those in the LMa and SMa groups in each treatment. The bacterial communities in the Mia and P particle sizes in each treatment group were more complex. Therefore, coinoculation of AM fungi and rhizobia stimulated atrazine dissipation by changing the atrazine-degrading bacterial community, and the response of the atrazine-degrading bacterial community to each aggregate size varied depending on its distinct soil physicochemical properties.

Beyond genome: Advanced omics progress of Panax ginseng.

Ginseng is a perennial herb of the genus Panax in the family Araliaceae as one of the most important traditional medicine. Genomic studies of ginseng assist in the systematic discovery of genes related to bioactive ginsenosides biosynthesis and resistance to stress, which are of great significance in the conservation of genetic resources and variety improvement. The transcriptome reflects the difference and consistency of gene expression, and transcriptomics studies of ginseng assist in screening ginseng differentially expressed genes to further explore the powerful gene source of ginseng. Protein is the ultimate bearer of ginseng life activities, and proteomic studies of ginseng assist in exploring the biosynthesis and regulation of secondary metabolites like ginsenosides and the molecular mechanism of ginseng adversity adaptation at the overall level. In this review, we summarize the current status of ginseng research in genomics, transcriptomics and proteomics, respectively. We also discuss and look forward to the development of ginseng genome allele mapping, ginseng spatiotemporal, single-cell transcriptome, as well as ginseng post-translational modification proteome. We hope that this review will contribute to the in-depth study of ginseng and provide a reference for future analysis of ginseng from a systems biology perspective.

Regional decreases of cortical thickness in major depressive disorder and their correlation with illness duration: a case-control study.

Background: Alterations in brain structure and function in major depressive disorder (MDD) have been identified in a number of studies, but findings regarding cortical thickness were various and inconsistent. Our current study aims to explore the differences in cortical thickness between individuals with MDD and healthy controls (HC) in a Chinese population. Methods: We investigated T1-weighted brain magnetic resonance imaging data from 61 participants (31 MDD and 30 HC). The cortical thickness between the two groups and analyzed correlations between cortical thickness and demographic variables in the MDD group for regions with significant between-group differences were conducted. Results: Compared with the HC group, patients with MDD had significantly decreased cortical thickness, in left pars triangularis, left pars orbitalis, left rostral middle frontal gyrus, left supramarginal gyrus, right parahippocampal gyrus, right lingual gyrus, right fusiform and right inferior parietal gyrus. The cortical thickness of left rostral middle frontal gyrus was negatively correlated (r = -0.47, p = 0.028) with the illness duration in patients with MDD. Conclusion: Our study distinguished that cortical thickness decreases in numerous brain regions both in the left and right hemisphere in individuals with MDD, and the negative correlation between the cortical thickness of left rostral middle frontal gyrus illness duration. Our current findings are valuable in providing neural markers to identify MDD and understanding the potential pathophysiology of mood disorders.

MOMS: A pipeline for scaffolding using multi-optical maps.

Here, we report a new multi-optical maps scaffolder (MOMS) aiming at utilizing complementary information among optical maps labelled by distinct enzymes. This pipeline was designed for data structure organization, scaffolding by path traversal, gap-filling and molecule reuse of optical maps. Our testing showed that this pipeline has uncapped enzyme tolerance in scaffolding. This means that there are no inbuilt limits as to the number of maps generated by different enzymes that can be utilized by MOMS. For the genome assembly of the human GM12878 cell line, MOMS significantly improved the contiguity and completeness with an up to 144-fold increase of scaffold N50 compared with initial assemblies. Benchmarking on the genomes of human and O. sativa showed that MOMS is more effective and robust compared with other optical-map-based scaffolders. We believe this pipeline will contribute to high-fidelity chromosome assembly and chromosome-level evolutionary analysis.

White matter microstructure changes in adults with major depressive disorder: evidence from diffusion magnetic resonance imaging.

BACKGROUND: Major depressive disorder (MDD) is a serious psychiatric disorder marked by low mood and anhedonia. Understanding the neural mechanism of MDD is essential for the treatment of depression. White matter fibres, connecting different computational units in the brain, have an important effect on brain function; however, the mechanism of white matter fibre abnormality in MDD is still unclear. AIMS: Our study expected to find white matter abnormalities associated with the frontal lobe and hippocampus in individuals with MDD. METHOD: Using diffusion tensor imaging data and tract-based spatial statistics, we investigated the microstructural differences in white matter fibre tracts between 30 adults with MDD compared with 31 healthy controls, and calculated the association between MDD-related microstructural changes and illness duration. RESULTS: It was found that patients with MDD showed reduced fractional anisotropy in the genu and body of the corpus callosum, right corona radiata and part of the thalamic radiations, suggesting lower fibrous myelination levels in these regions; the decreased fractional anisotropy in these regions was associated with longer illness duration. CONCLUSIONS: Our results suggest that MDD may be associated with microstructural damage of key fibre tracts, which could provide insights into the understanding and treatment of MDD.

Neural activity in adults with major depressive disorder differs from that in healthy individuals: A resting-state functional magnetic resonance imaging study.

Objective: Currently, findings regarding resting-state functional magnetic resonance imaging studies of major depressive disorder (MDD) are inconsistent. In contrast to the previously used a priori seed-based functional connectivity analyses, this study employed whole-brain exploratory analyses and aimed to explore neural activity patterns in Chinese adults with MDD. Materials and methods: Specifically, this study examined the amplitude of low-frequency fluctuations within the whole brain and adopted a large-scale brain network template to explore the core dysfunctional brain regions in individuals with MDD. Results: Overall, 32 individuals with MDD and 32 healthy controls were evaluated. Compared to healthy controls, individuals with MDD showed more profound alterations in the amplitude of low-frequency fluctuations in the temporolimbic affective circuit (e.g., middle temporal gyrus and parahippocampus) and default mode network (e.g., precuneus and thalamus). Moreover, functional connectivity between the left mid-insula and parietal regions within the sensorimotor network was weaker in individuals with MDD than in healthy controls. Conclusion: In conclusion, the neural characteristics of MDD correspond to cognitive deficits in self-referential processing and emotional processing and are related to a risk of sensory disorders or psychomotor retardation. These findings present neural markers that may be used to identify MDD, contributing to clinical diagnosis.

The Gut Microbial Co-Abundance Gene Groups (CAGs) Differentially Respond to the Flavor (Yao-Wei) of Chinese Materia Medica.

The property theory is a unique principle instructing traditional Chinese doctors to prescribe proper medicines against diseases. As an essential part of it, the five-flavor theory catalogs various Chinese materia medicas (CMMs) into five flavors (sweet, bitter, sour, salty, and pungent) based on their taste and medical functions. Although CMM has been successfully applied in China for thousands of years, it is still a big challenge to interpret CMM flavor via modern biomarkers, further deepening its elusiveness. Herein, to identify the correlation between gut microbiota and CMM flavor, we selected 14 CMMs with different flavors to prepare their aqueous extracts, quantified the contained major chemical components, and then performed full-length 16S rRNA sequencing to analyze the gut microbiota of C57BL/6 mice administrated with CMM extracts. We found that flavones, alkaloids, and saponins were the richest components for sweet-, bitter-, and pungent-flavored CMMs, respectively. Medicines with merged flavors (bitter-pungent and sweet-pungent) displayed mixed profiles of components. According to gut microbial analysis, modulation of CMMs belonging to the same flavor on the taxonomic classification was inconsistent to an extent, while the functional sets of gut microbiota, co-abundance gene groups (CAGs), strongly and differentially responded to distinct flavors. Moreover, these correlations were in line with their pharmacological actions. Therefore, the gut microbial functional sets (CAGs) could act as the possible indicator to reflect CMM flavor, rather than the composition of microbial community.

Preparation, optimization, and in vivo evaluation of an inhaled solution of total saponins of Panax notoginseng and its protective effect against idiopathic pulmonary fibrosis.

Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive pulmonary disease that can cause fibrotic remodeling of the surrounding lung, thus leading to respiratory failure. Although IPF is the most common form of idiopathic interstitial pneumonia, the precise mechanisms underlying this condition remain unknown. In this study, we used total saponins of Panax notoginseng inhalation solution (TIS) to induce idiopathic bleomycin-induced pulmonary fibrosis in rats. The uniformity of delivery dose was investigated by analyzing the aerodynamic particle size distribution and drug stability. The potential of hydrogen potential of hydrogen (pH) of the inhalation solution was 7.0 and the solvent 0.9% NaCl solution, thus meeting physiological requirements for pulmonary drug administration. The delivery rate was 1.94 ± 0.16 mg·min-1 and the total dose was 17.40 ± 0.04 mg. TIS was composed of five key components: notoginsenoside R1, ginsenosides Rg1, ginsenosides Re, ginsenosides Rb1, and ginsenosides Rd. The mass median aerodynamic diameter (MMAD) for these five components were 3.62 ± 0.05 µm, 3.62 ± 0.06 µm, 3.65 ± 0.10 µm, 3.62 ± 0.06 µm, and 3.61 ± 0.05 µm, respectively. Fine particle fraction (FPF) was 66.24 ± 0.73%, 66.20 ± 0.89%, 66.07 ± 1.42%, 66.18 ± 0.79%, and 66.29 ± 0.70%, respectively. The MMAD for inhalation solutions needs to be 1-5 µm, which indicates that the components of TIS are suitable for inhalation. It is important to control the particle size of targeted drugs to ensure that the drug is delivered to the appropriate target tissue. In vitro experiments indicated that TIS exhibited high rates of deposition in lung tissue, thus indicating that pulmonary delivery systems may represent a good therapeutic option for patients.

pH-Responsive Artesunate Polymer Prodrugs with Enhanced Ablation Effect on Rodent Xenograft Colon Cancer.

PURPOSE: In this study, pH-sensitive poly(2-ethyl-2-oxazoline)-poly(lactic acid)-poly(ß-amino ester) (PEOz-PLA-PBAE) triblock copolymers were synthesized and were conjugated with an antimalaria drug artesunate (ART), for inhibition of a colon cancer xenograft model. METHODS: The as-prepared polymer prodrugs are tended to self-assemble into polymeric micelles in aqueous milieu, with PEOz segment as hydrophilic shell and PLA-PBAE segment as hydrophobic core. RESULTS: The pH sensitivity of the as-prepared copolymers was confirmed by acid-base titration with pKb values around 6.5. The drug-conjugated polymer micelles showed high stability for at least 96 h in PBS and 37°C, respectively. The as-prepared copolymer prodrugs showed high drug loading content, with 9.57%±1.24% of drug loading for PEOz-PLA-PBAE-ART4. The conjugated ART could be released in a sustained and pH-dependent manner, with 92% of released drug at pH 6.0 and 57% of drug released at pH 7.4, respectively. In addition, in vitro experiments showed higher inhibitory effect of the prodrugs on rodent CT-26 cells than that of free ART. Animal studies also demonstrated the enhanced inhibitory efficacy of PEOz-PLA-PBAE-ART2 micelles on the growth of rodent xenograft tumor. CONCLUSION: The pH-responsive artesunate polymer prodrugs are promising candidates for colon cancer adjuvant therapy.

Inactivation of endothelial ZEB1 impedes tumor progression and sensitizes tumors to conventional therapies.

Current antiangiogenic therapy is limited by its cytostatic property, scarce drug delivery to the tumor, and side toxicity. To address these limitations, we unveiled the role of ZEB1, a tumor endothelium-enriched zinc-finger transcription factor, during tumor progression. We discovered that the patients who had lung adenocarcinomas with high ZEB1 expression in tumor endothelium had increased prevalence of metastases and markedly reduced overall survival after the diagnosis of lung cancer. Endothelial ZEB1 deletion in tumor-bearing mice diminished tumor angiogenesis while eliciting persistent tumor vascular normalization by epigenetically repressing TGF-ß signaling. This consequently led to improved blood and oxygen perfusion, enhanced chemotherapy delivery and immune effector cell infiltration, and reduced tumor growth and metastasis. Moreover, targeting vascular ZEB1 remarkably potentiated the anticancer activity of nontoxic low-dose cisplatin. Treatment with low-dose anti-programmed cell death protein 1 (anti-PD-1) antibody elicited tumor regression and markedly extended survival in ZEB1-deleted mice, conferring long-term protective anticancer immunity. Collectively, we demonstrated that inactivation of endothelial ZEB1 may offer alternative opportunities for cancer therapy with minimal side effects. Targeting endothelium-derived ZEB1 in combination with conventional chemotherapy or immune checkpoint blockade therapy may yield a potent and superior anticancer effect.

[Preparation of docetaxel-loaded nanomicelles and their anti-Lewis lung cancer effect in vitro].

Docetaxel-loaded nanomicelles were prepared in this study to improve the solubility and tumor targeting effect of docetaxel(DTX),and further evaluate their anticancer effects in vitro. PBAE-DTX nanomicelles were prepared by film-hydration method with amphiphilic block copolymer polyethyleneglycol methoxy-polylactide(PELA) and pH sensitive triblock copolymer polyethyleneglycol methoxy-polylactide-poly-ß-aminoester(PBAE) were used respectively to prepare PELA-DTX nanomicelles and PBAE-DTX nanomicelles. The nanomicelles were characterized by physicochemical properties and the activity of mice Lewis lung cancer cells was studied. The results of particle size measurement showed that the blank micelles and drug-loaded micelles had similar particle sizes, ranging from 10 to 100 nm. The particle size of PBAE micelles was changed under weak acidic conditions, with good pH response. The encapsulation efficiency of the above two types of DTX-loaded nanomicelles determined by HPLC was(93.8±1.70)% and(87.2±4.10)%, and the drug loading amount was(5.3±0.10)% and(4.9±0.05)%,respectively. Furthermore,the DTX micelles also showed significant inhibitory effects on Lewis lung cancer cells by MTT assay, and pH-sensitive PBAE-DTX showed better cytotoxicity. The results of flow cytometry indicated that,the apoptosis rate of lung cancer Lewis cells was(20.72±1.47)%,(29.71±2.38)%,and(40.91±1.90)%(P<0.05) at 48 h after treatment in DTX,PELA-DTX,and PBAE-DTX groups. The results showed that different docetaxel preparations could promote the apoptosis of Lewis cells, and PBAE-DTX had stronger apoptotic-promoting effect. The pH-sensitive DTX-loaded micelles are promising candidates in developing stimuli triggered drug delivery systems in acidic tumor micro-environments with improved inhibitory effects of tumor growth on Lewis lung cancer.

Simultaneous determination of seven flavonoids, two phenolic acids and two cholesterines in Tanreqing injection by UHPLC-MS/MS.

A new ultra-high performance liquid chromatography combined with triple quadrupole mass spectrometry was developed to evaluate the quality of Tanreqing injection. Seven flavonoids (Rutin, Baicalin, Scutellarin, Chrysin-7-O-Beta-d-glucoronide, Oroxylin A-7-O-ß-d-glucoronide, Wogonin, Luteolin-7-O-glucoside), two phenolic acids (Chlorogenic acid, Caffeic acid) and two cholesterines (Ursodeoxycholic acid, Chenodeoxycholic acid) in Tanreqing injection could be measured simultaneously. For the determination of the eleven compounds, the conditions were set as follows: The mobile phase was a gradient of 0.1% aqueous formic acid solution (A) and acetonitrile (B); the flow rate was 0.2 mL min-1, the column was Acquity UPLC HSS T3 column (2.1 mm × 100 mm, 1.8 µm); and the multiple-reaction monitoring (MRM) with a negative electro spray ionization interface (ESI-) was selected. Within the test ranges, all the standard regression curves showed excellent linear regression (r > 0.99). In terms of (relative standard deviation) RSDs, the precision, repeatability and stability of the eleven compounds were all lower than 3%. The recovery rates of Tanreqing injection and the RSD were 97.8-103.7% and 0.4%-2.0%, respectively. The RSD value was in accordance with the requirements of less than 3.0%. This method has been successfully used in the analysis of Tanreqing injection. In summary, a fast, accurate and reliable UPLC-ESI--MS/MS method was successfully developed for the simultaneous detection of the eleven major active ingredients with different chemical structures in Tanreqing injection, and can be used for the quality control of Tanreqing injection as well.

[Freeze drying process optimization of ginger juice-adjuvant for Chinese materia medica processing and stability of freeze-dried ginger juice powder].

Ginger juice， a commonly used adjuvant for Chinese materia medica， is applied in processing of multiple Chinese herbal decoction pieces. Because of the raw materials and preparation process of ginger juice， it is difficult to be preserved for a long time， and the dosage of ginger juice in the processing can not be determined base on its content of main compositions. Ginger juice from different sources is hard to achieve consistent effect during the processing of traditional Chinese herbal decoction pieces. Based on the previous studies， the freeze drying of ginger juice under different shelf temperatures and vacuum degrees were studied， and the optimized freeze drying condition of ginger juice was determined. The content determination method for 6-gingerol， 8-gingerol， 10-gingerol and 6-shagaol in ginger juice and redissolved ginger juice was established. The content changes of 6-gingerol， 8-gingerol， 10-gingerol， 6-gingerol， 6-shagaol， volatile oil and total phenol were studied through the drying process and 30 days preservation period. The results showed that the freeze drying time of ginger juice was shortened after process optimization; the compositions basically remained unchanged after freeze drying， and there was no significant changes in the total phenol content and gingerol content， but the volatile oil content was significantly decreased(P<0.05). Within 30 days， the contents of gingerol， total phenol， and volatile oil were on the decline as a whole. This study has preliminarily proved the feasibility of freeze-drying process of ginger juice as an adjuvant for Chinese medicine processing.

The Effect of Ginger Juice Processing on the Chemical Profiles of Rhizoma coptidis.

Rhizoma coptidis (RC) has been used as an herbal medicine in China for over one thousand years, and it was subjected to specific processing before use as materia medica. Processing is a pharmaceutical technique that aims to enhance the efficacy and/or reduce the toxicity of crude drugs according to traditional Chinese medicine theory. In this study, the chemical profiles of RC, ginger juice processed RC (GRC), and water processed RC (WRC) was determined to reveal the mechanism of processing of RC. UPLC-QTOF-MS analysis of methanol extract of RC, GRC, and WRC has been conducted to investigate the effect of processing on the composition of RC. HPLC-PDA was used to determine the variance of total alkaloids and seven alkaloids of RC during the processing. The volatiles of RC, GRC and ginger juice were separated by distillation, the change of volatiles content was recorded and analyzed, and the qualitative analysis of the volatiles was carried out using GC-MS. The microstructures of RC, GRC and WRC were observed using a light microscope. Results showed that ginger juice/water processing had limited influence on the composition of RC's methanol extract, but significant influence on the content of some alkaloids in RC. Ginger juice processing significantly increased (p < 0.05) the volatiles content of RC and changed the volatiles composition obviously. Processing also had an influence on the microstructure of RC. This research comprehensively revealed the mechanism of ginger juice processing of RC.

EjODO1, a MYB Transcription Factor, Regulating Lignin Biosynthesis in Developing Loquat (Eriobotrya japonica) Fruit.

Lignin is important for plant secondary cell wall formation and participates in resistance to various biotic and abiotic stresses. Loquat undergoes lignification not only in vegetative tissues but also in flesh of postharvest fruit, which adversely affects consumer acceptance. Thus, researches on lignin biosynthesis and regulation are important to understand loquat fruit lignification. In loquat, a gene encoding an enzyme in the lignin biosynthesis pathway, Ej4CL1, was reported to be regulated by transcription factors, including EjMYB1, EjMYB2, EjMYB8, and EjAP2-1, knowledge of this process is still limited. With the aim of identifying novel transcriptional factors controlling lignin biosynthesis in loquat, the promoter of Ej4CL1 was utilized to screen a cDNA library by yeast one hybrid assay. A novel R2R3 MYB, named EjODO1, was identified. Real-time PCR analyses indicated that EjODO1 is highly expressed in lignified stems and roots. During fruit development, expression of EjODO1 decreased along with the reduction of lignin content and became undetectable in mature ripe fruit. Thus, EjODO1 is likely to be involved in lignification of vegetative organs and early fruit development but not in mature fruit or postharvest lignification. Dual-luciferase assay indicated that EjODO1 could trans-activate promoters of lignin biosynthesis genes, such as EjPAL1, Ej4CL1, and Ej4CL5 and transient overexpression of EjODO1 triggered lignin biosynthesis. These results indicate a role for EjODO1 in regulating lignin biosynthesis in loquat which is different from the previously characterized transcription factors.

Self-Assembled Thermoresponsive Nanogels Prepared by Reverse Micelle → Positive Micelle Method for Ophthalmic Delivery of Muscone, a Poorly Water-Soluble Drug.

This study aimed to design a nanocarrier ophthalmic delivery system of muscone, a poorly water-soluble drug. The muscone thermoresponsive nanogels were self-assembled by reverse micelle → positive micelle method. Muscone was demonstrated to have uniform narrow particle size distribution in nanogel by the dynamic light scattering test. The developed nanocomposite hydrogel had a high muscone loading, and the rheology results showed that the phase transition temperature was 34.05°C. Thixotropy test indicated that the nanogel was able to resist the blinking of eyes because of the thixotropy recovery time, which is <5 s. Compared with muscone eye drops, muscone nanogels showed longer retention time on the corneal surface using fluorescent labeling technology and produced a 3.4-fold increase in apparent permeability coefficients (Papp). Draize testing showed that the developed nanogel caused no eye irritation. In vivo pharmacokinetic study indicated that the nanogel could significantly increase the bioavailability of muscone after administration compared with eye drops. These results indicate that self-assembled thermoresponsive nanogel prepared by reverse micelle → positive micelle method has potential for the ophthalmic delivery of poorly water-soluble drugs.

[Preparation and evaluation of Shedan in situ forming gel based on ocular characteristics].

To develop an ophthalmic preparation of Shedan, an in situ forming gel was prepared with the formulation containing 18% of poloxamer 407 and 5% of poloxamer 188 by response surface designs plus central composite designs. The rheology results showed that LVE range gamma should limited within 0.5%, Shedan high-frequency region, and the thixotropy recovery time is less than 5 seconds. The phase transition temperature was 33.25 °C according to curve of storage modulus and loss modulus determined by temperature scanning. Surface tension and osmometer of it determined by surface tension meter and dew point osmometer were 36.43 mN · m(-1), and 320.6 mOsm · kg(-1), respectively. Fluorescein sodium was selected as the marker to monitor the corneal residence time, and the results showed that Shedan gel could prolong drug residence for 180 min. In line with zero-order kinetics, releases of muscone and salvianolic acid B in vitro depends on gels erosion. The results of rabbit ocular irritation experiments suggested that Shedan in situ forming gel was biocompatible and nonirritant. In conclusion, a novel Shedan in situ forming gel was developed and characterized for potential drug treatment of retinal vein occlusion.

[Study on preparation of Dange Mingmu in-situ forming eye gel].

OBJECTIVE: To prepare Dange Mingmu in-situ forming eye gel. METHOD: The viscosity of Dange Mingmu in-situ forming eye gel was tested by adopting poloxamer 407 and 188 as thermosensitive materials, and optimizing by uniform design. Drug release in vitro was studied using a novel membraneless model. Eye irritation experiments were performed with rabbits. The duration of residence time in rabbit eyes was observed using fluorescence tracer method. RESULT: The gelation temperature of in-situ thermosensitive gel was lowered as the P407 concentration increased. In a certain range, the gelation temperature slowly increased with the increase of P188's concentration, and the effect of P407 was greater than that of P188. The optimized concentration of P407/P188 was 19%/1%. Based the adjusted concentration, Dange Mingmu in-situ forming eye gel. was converted into freely flowing liquid below 26.9 degrees C and became gel at 34.5 degrees C after being diluted with STF. In line with zero-order kinetics, drug release in vitro depends on gel erosion. The residence time on the surface of eyes was proved to be relatively long was and nonirritant. CONCLUSION: Uniform design is available for optimizing the formulation of thermosensitive gel for eye. The gel satisfies the requirement for ophthalmic application, and is expected to be applied in clinical practice in the future.

Ultrafast dynamics of nonequilibrium resonance energy transfer and probing globular protein flexibility of myoglobin.

Protein structural plasticity is critical to many biological activities and accurate determination of its temporal and spatial fluctuations is challenging and difficult. Here, we report our extensive characterization of global flexibility of a globular heme protein of myoglobin using resonance energy transfer as a molecular ruler. With site-directed mutagenesis, we use a tryptophan scan to examine local structural fluctuations from B to H helices utilizing 10 tryptophan-heme energy transfer pairs with femtosecond resolution. We observed ultrafast resonance energy transfer dynamics by following a nearly single exponential behavior in 10-100 ps, strongly indicating that the globular structure of myoglobin is relatively rigid, with no observable static or slow dynamic conformational heterogeneity. The observation is against our molecular dynamics simulations, which show large local fluctuations and give multiple exponential energy transfer behaviors, suggesting too flexible of the global structure and thus raising a serious issue of the force fields used in simulations. Finally, these ultrafast energy transfer dynamics all occur on the similar time scales of local environmental relaxations (solvation), leading to nonexponential processes caused by energy relaxations, not structural fluctuations. Our analyses of such processes reveal an intrinsic compressed- and/or stretched-exponential behaviors and elucidate the nature of inherent nonequilibrium of ultrafast resonance energy transfer in proteins. This new concept of compressed nonequilibrium transfer dynamics should be applied to all protein studies by time-resolved Förster resonance energy transfer (FRET).

[Preparation and characterization of non-ionic surfactant vesicle of cantharidin].

OBJECTIVE: To study the preparation of cantharidin entrapped non-ionic surfactant vesicle (noisome)and evaluate its quality. METHOD: The niosome loaded with cantharidin was prepared using injection method by non-ionic surfactants as the carrier. An centrifugation separation method and HPLC analysis method of the cantharidin were established to detect the entrapment efficiency. The optimum preparation technology was established by a orthogonal experiment. The morphology, and particle size were studied to evaluate the preparation. RESULT: The average size of niosomes were (209. 8 +/- 0.5) nm. The entrapment efficiency of the CTD-NS was (27.5% +/- 2.0%) and Zeta potential was (41.5 +/- 0.65) mV. CONCLUSION: The preparation of cantharidin noisome by TweenA and SpanB is practicable and successful. These experiments can be the basement of developing targeting drug delivery system.