An optofluidic system for the concentration gradient screening of oocyte protection drugs.

Oocytes protective drug screening is essential for the treatment of reproductive diseases. However, few studies construct the oocyte in vitro drug screening microfluidic systems because of their enormous size, scarcity, and sensitivity to the culture environment. Here, we present an optofluidic system for oocyte drug screening and state analysis. The system consists of two parts: an open-top drug screening microfluidic chip and an optical Fourier filter analysis part. The open-top microfluidic chip anchors single oocyte with hydrogel and allows nutrient and gas environment updating which is essential for oocyte culturing. The optical filter analysis part is used to accurately analyse the status of oocytes. Based on this system, we found that fluorene-9-bisphenol (BHPF) damaged the oocyte spindle in a dose-dependent manner, a high dose of melatonin (10-3 M) effectively reduces the percentage of abnormally arranged chromosomes of oocytes exposed to 40 µM BHPF. This optofluidic system shows great promise for the culture of oocytes and demonstrates the robust ability for convenient multi-concentration oocytes drug screening. This technology may benefit further biomedicine and reproductive toxicology applications in the lab on a chip community.

Transcriptome Analysis Reveals the Biocontrol Mechanism of Endophytic Bacterium AM201, Rhodococcus sp., against Root Rot Disease of Atractylodes macrocephala.

Atractylodes macrocephala Koidz (AMK) is a perennial herb from the plant family Asteraceae (formerly Compositae). This herb is mainly distributed in mountainous wetlands in Zhejiang, Sichuan, Yunnan, and Hunan provinces of China. Its medicinal production and quality, however, are severely impacted by root rot disease. In our previous study, endophytic bacterium designated AM201 exerted a high biocontrol effect on the root rot disease of AMK. However, the molecular mechanisms underlying this effect remain unclear. In this study, the identity of strain AM201 as Rhodococcus sp. was determined through analysis of its morphology, physiological and biochemical characteristics, as well as 16S rDNA sequencing. Subsequently, we performed transcriptome sequencing and bioinformatics analysis to compare and analyze the transcriptome profiles of root tissues from two groups: AM201 (AMK seedlings inoculated with Fusarium solani [FS] and AM201) and FS (AMK seedlings inoculated with FS alone). We also conducted morphological, physiological, biochemical, and molecular identification analyses for the AM201 strain. We obtained 1,560 differentially expressed genes, including 187 upregulated genes and 1,373 downregulated genes. We screened six key genes (GOLS2, CIPK25, ABI2, egID, PG1, and pgxB) involved in the resistance of AM201 against AMK root rot disease. These genes play a critical role in reactive oxygen species (ROS) clearance, Ca2+ signal transduction, abscisic acid signal inhibition, plant root growth, and plant cell wall defense. The strain AM201 was identified as Rhodococcus sp. based on its morphological characteristics, physiological and biochemical properties, and 16S rDNA sequencing results. The findings of this study could enable to prevent and control root rot disease in AMK and could offer theoretical guidance for the agricultural production of other medicinal herbs.

Acoustic quasi-periodic bioassembly based diverse stem cell arrangements for differentiation guidance.

Arrangement patterns and geometric cues have been demonstrated to influence cell function and fate, which calls for efficient and versatile cell patterning techniques. Despite constant achievements that mainly focus on individual cells and uniform cell patterns, simultaneously constructing cellular arrangements with diverse patterns and positional relationships in a flexible and contact-free manner remains a challenge. Here, stem cell arrangements possessing multiple geometries and structures are proposed based on powerful and diverse pattern-building capabilities of quasi-periodic acoustic fields, with advantages of rich patterns and structures and flexibility in structure modulation. Eight-fold waves' interference produces regular potentials that result in higher rotational symmetry and more complex arrangement of geometric units. Moreover, through flexible modulation of the phase relations among these wave vectors, a wide variety of cellular pattern units are arranged in this potential, such as circular-, triangular- and square-shape, simultaneously. It is proved that these diverse cellular patterns conveniently build human mesenchymal stem cell (hMSC) models, for research on the effect of cellular arrangement on stem cell differentiation. This work fills the gap of acoustic cell patterning in quasi-periodic patterns and shows promising potential in tissue engineering and regenerative medicine.

Convenient tumor 3D spheroid arrays manufacturing via acoustic excited bubbles for in situ drug screening.

The quick and convenient fabrication of in vitro tumor spheroids models has been pursued for clinical drug discovery and personalized therapy. Here, uniform three-dimensional (3D) tumor spheroids are quickly constructed by acoustically excited bubble arrays in a microfluidic chip and performed drug response testing in situ. In detail, bubble oscillation excited by acoustic waves induces second radiation force, resulting in the cells rotating and aggregating into tumor spheroids, which obtain controllable sizes ranging from 30 to 300 µm. These spherical tumor models are located in microfluidic networks, where drug solutions with gradient concentrations are generated from 0 to 18 mg mL-1, so that the cell spheroids response to drugs can be monitored conveniently and efficiently. This one-step tumor spheroids manufacturing method significantly reduces the model construction time to less than 15 s and increases efficiency by eliminating additional transfer processes. These significant advantages of convenience and high-throughput manufacturing make the tumor models promising for use in tumor treatment and point-of-care diagnosis.

On-demand liquid microlens arrays by non-contact relocation of inhomogeneous fluids in acoustic fields.

Microlens arrays (MLAs) are key micro-optical components that possess a high degree of parallelism and ease of integration. However, rapid and low-cost fabrication of MLAs with flexible focusing remains a challenge. Herein, liquid MLAs with dynamic tunability are presented using non-contact acoustic relocation of inhomogeneous fluids. By designing ring-shaped acoustic pressure node (PN) arrays, the denser fluid of miscible liquids is relocated to PNs, and liquid MLAs with ideal morphology are obtained. The experimental results demonstrate that the liquid MLAs possess a powerful reconfigurability with long-term stability and sharp imaging that can conveniently switch between the on and off state and can dynamically magnify by simply adjusting the acoustic amplitude. Moreover, the high biocompatibility inherited from liquids accompanied by the acoustic treatment allows cells to be within working distance of the MLAs without immersion, as would be required for a solid lens. This innovative liquid MLA is inexpensive to manufacture and possesses continuous focus, fast response, and satisfactory bioaffinity, and thus offers promising potential for microfluidic adaptive imaging and biomedical sensing, especially for live cell imaging.

The phytochemical properties, pharmacological effects and traditional uses of Actinidia eriantha Benth.: A review.

Actinidia eriantha Benth. (Called Maohuamihoutao in China) is a plant that has been utilized as a heat-clearing drug in She ethnic minority group for a long time in China. Specifically, it has been involved in the treatment of stomach cancer, colon cancer, cirrhosis with ascites, chronic hepatitis, leukemia, rectal prolapse, hernia and uterine prolapse. Pharmacological research provides partial evidence for the traditional use of A. eriantha and might have demonstrated the folk utilization of A. eriantha to combat many cancers. Crude extracts and relatively pure components of A. eriantha possess a variety of pharmacological activities, including anti-cancer, immunoregulatory, anti-angiogenic, neuroprotective, anti-inflammatory, and antioxidant activities. In addition, over 104 chemical substances have been determined from A. eriantha, involving terpenoids, alcohols, phenolics, aldehydes, organic acids, flavonoids glycosides, ketones, and glucoside. The existing literature reveals that a large proportion of the therapeutic effects of A. eriantha were rendered by the polysaccharides. However, the mechanisms of action and the structure-function correlations of these compounds, as well as the synergistic and antagonistic effects between them, need to be investigated further. Therefore, we propose that future studies on A. eriantha should focus on comprehensively assessing its medicinal quality, exploring its multi-target nature using network pharmacology approaches, and evaluating its long-term toxicity and efficacy in vivo.

Dynamics of vaginal microbiome in female beagles at different ages.

Age-related changes in human vaginal microbiota composition have been reported, and such changes might be influenced by humidity, external stimuli, hormone levels, drug to use, and other factors. However, there is no report about the vaginal microbiota composition of female beagles at different ages. To investigate the effects of aging on the vaginal microbiota independent of other effects, we analyzed the vaginal microbiomes of 23 beagles at a wide range of ages from 1 year to 7 years old (except the 3rd year), 1-2 y were categorized into youth stage (YS), 4-5 y were categorized into middle stage (MS), and 6-7 y were categorized into elderly stage (ES) based on age. Samples were collected by scraping the vaginal mucosa of YS (n = 8), MS (n = 5) and ES (n = 10), and analyzed by 16S-rRNA gene high-throughput-sequencing. The diversity of the vaginal microbiome in female beagles was found to continuously change with age. We also found associations between age and specific members and functions of the vaginal microbiome. The metabolism of terpenoids and polyketide and the cell motility are significantly enhanced with age. Our results suggest that the proportion of Tenericutes might be a biomarker which could distinguish between YS and others.

Research on Preparation Methods of Carbon Nanomaterials Based on Self-Assembly of Carbon Quantum Dots.

Here, based on self-assembly of carbon quantum dots (CDs), an innovative method to prepare nanomaterials under the action of a metal catalyst was presented. CDs were synthesized by a one-step hydrothermal method with citric acid (CA) as the carbon source, ethylenediamine (EDA) as the passivator and FeSO4•7H2O as the pre-catalyst. In the experiment, it was found that the nano-carbon films with a graphene-like structure were formed on the surface of the solution. The structure of the films was studied by high-resolution transmission electron microscopy (HRTEM), Fourier transform infrared (FT-IR), etc. The results demonstrated that the films were formed by the self-assembly of CDs under the action of the gas-liquid interface template and the metal catalyst. Meanwhile, the electrochemical performance of the films was evaluated by linear cyclic voltammetry (CV) and galvanostatic charge discharge (GOD) tests. In addition, the bulk solution could be further reacted and self-assembled by reflux to form a bifunctional magnetic-fluorescent composite material. Characterizations such as X-ray diffractometer (XRD), fluorescence spectra (FL), vibrating sample magnetometer (VSM), etc. revealed that it was a composite of superparamagnetic γ-Fe2O3 and CDs. The results showed that self-assembly of CDs is a novel and effective method for preparing new carbon nanomaterials.

Touchable cell biophysics property recognition platforms enable multifunctional blood smart health care.

As a crucial biophysical property, red blood cell (RBC) deformability is pathologically altered in numerous disease states, and biochemical and structural changes occur over time in stored samples of otherwise normal RBCs. However, there is still a gap in applying it further to point-of-care blood devices due to the large external equipment (high-resolution microscope and microfluidic pump), associated operational difficulties, and professional analysis. Herein, we revolutionarily propose a smart optofluidic system to provide a differential diagnosis for blood testing via precise cell biophysics property recognition both mechanically and morphologically. Deformation of the RBC population is caused by pressing the hydrogel via an integrated mechanical transfer device. The biophysical properties of the cell population are obtained by the designed smartphone algorithm. Artificial intelligence-based modeling of cell biophysics properties related to blood diseases and quality was developed for online testing. We currently achieve 100% diagnostic accuracy for five typical clinical blood diseases (90 megaloblastic anemia, 78 myelofibrosis, 84 iron deficiency anemia, 48 thrombotic thrombocytopenic purpura, and 48 thalassemias) via real-world prospective implementation; furthermore, personalized blood quality (for transfusion in cardiac surgery) monitoring is achieved with an accuracy of 96.9%. This work suggests a potential basis for next-generation blood smart health care devices.

A Facile and Scalable Fabrication Procedure for Thin-Film Composite Membranes: Integration of Phase Inversion and Interfacial Polymerization.

Conventional dense thin-film composite (TFC) membranes evince a universally low water permeability, the increase of which typically relies on introducing additional transport channels based on intricate steps within a membrane preparation process. In this study, we reported a novel and simplified procedure for the fabrication of high-performance TFC membranes. Specifically, the dissolution of aqueous monomers in the casting solution was utilized for the following interfacial polymerization (IP). Since the monomers diffused to the water bath during phase inversion, the control of precipitation time enabled an effective regulation of the monomer concentration in the formed polymeric substrates, where the IP reaction was initiated by the addition of the organic phase. The entire and uniform embedment of aqueous monomers inside the substrates contributed to the formation of ultrathin and smooth selective layers. An excellent separation performance (i.e., water permeability: 34.7 L m-2 h-1 bar-1; Na2SO4 rejection: ∼96%) could be attained using two types of aqueous monomers (i.e., piperazine and ß-cyclodextrin), demonstrating the effectiveness and universality of this method. Compared to the conventional immersion-based process, this novel procedure shows distinct advantages in reducing monomer usage, shortening the production cycle, and achieving a more superior membrane performance, which is highly promising for large-scale membrane manufacture.

Sensitively and Selectively Detect Biothiols by Using Fluorescence Method and Resonance Light Scattering Technique Simultaneously.

In this paper, we designed a new quantitative and qualitive detection method for biothiols by using fluorescence method and resonance light scattering (RLS) technique. Nitrogen doped carbon quantum dots (C/N-dots) were obtained from tartaric acid and ethylenediamine by hydrothermal method, and then their morphology and optical properties were characterized by different techniques. A detection system consisting of C/N-dots and Ag+ complex was established. In this system, C/N-dots possessed the photoluminescent property and the Ag+ complex owned the RLS property, so, by combining the two luminescent properties to achieve complementary advantages, we could detect biothiols and solve the problem of distinguishing between Cys and GSH. Additionally, we optimized detecting conditions and investigated the detection mechanism of fluorescence quenching and RLS detecting. Results showed that the analytical response of fluorescence was linear in the range 0-140 µM and the detection limit (LOD) was calculated to be 6.6 µM for Cys, and the addition of GSH had no effect on fluorescence. RLS response ranges were 0-167 µM for Cys and 0-200 µM for GSH, with LOD down to 64 nM and 74 nM, respectively. Furthermore, the probe was successfully used for detecting Cys in fetal bovine serum (FBS) samples by fluorescence method, and also, by RLS technique, the content of GSH in FBS samples was detected.

[Response of bacterial community structures at No. 10 Spring in Urumqi to felt earthquakes].

OBJECTIVE: Our aim was to know response of spring bacteria and metabolic characteristics of sensitive bacteria to felt earthquake. METHODS: Water samples were collected from January 31 to December 31, 2012, during which period 5 felt earthquakes occurred and the epicenter was 100 kilometers away from the No. 10 Spring in Urumqi. We monitored the spring bacterial activities and function diversity changes from No. 10 Spring in Urumqi during the pre- and post-earthquake stages by using plate culture counting methods and BIOLOG GEN III bacteria plate. RESULTS: The spring bacterial numbers presented stochastic dynamic changes through the year. The culturable bacteria numbers and average well color development (AWCD ) of carbon source utilization of bacterial community were higher after the earthquake. Besides, there were some correlations with magnitude and epicenter distance of earthquake. The main carbon source utilization types of sensitive bacteria group for felt earthquake were sugar alcohol at the No. 10 Spring. CONCLUSION: The results indicated that the BIOLOG GEN III plate can be used for spring bacterial metabolism diversity research. Culturable bacteria numbers and carbon source utilization of bacterial communities showed some reflecting earthquake law.

[Reliability and validity of DSSI-23 scale in a rural elderly population].

OBJECTIVE: To determine the reliability and validity of the Duke Social Support Index (DSSI)-23 scale in a rural elderly population. METHODS: A stratified cluster random sampling strategy was adopted to select 702 rural participants aged 60 years or over in Guangyuan. Those with severe hearing disorders or mental problems were excluded. The reliability and validity of DSSI-23 scale were evaluated using internal consistency, split-half reliability,content validity, convergent validity, discriminant validity and structure validity. RESULTS: The DSSI-23 had a Cronbach's alpha efficient of 0.881 and 0.918 split-half reliability. Item-scale correlation coefficients exceed 0.35 (P<0.01), except for item three. Satisfactory (100%) convergent validity and discriminant validity were found. Confirmatory Factor Analysis (CFA) demonstrated unsatisfactory fit with the theoretical model of the scale: chi2/df = 6.884, CFI=0.807, GFI=0.850, RMSEA=0.092. CONCLUSION: DSSI-23 scale has good reliability in the elderly population, but unsatisfactory validity. Further adjustments of the scale are needed.

[Current situation and relevant risk factors on sexual violence in 18-34 year-olds men in Chengdu].

OBJECTIVE: To understand the current situation and relevant factors on sexual violence in 18-34 year-olds men in Chengdu, in order to provide data for setting up prevention and reduction programs on sexual violence and other related problems. METHODS: By multi-stage random sampling, young men who were 18-34 years old in Chengdu were interviewed and screened. Questionnaire and multivariate logistic regression models were used to assess risk factors on sexual violence. RESULTS: The overall prevalence of sexual violence was 17.8% , with factors as child exposure to parent violence/family conflict (P < 0.01), child physical abuse (P < 0.01), alcohol abuse (P < 0.01), anxiety (P < 0.01), disharmony marriage (divorce/widowed/separation) (P < 0.01), marriage status (P < 0.05), depression (P < 0.05) as risk factors of sexual violence. CONCLUSION: The status on sexual violence among young men in Chengdu should not be overlooked and interventions for high-risk populations should be taken urqently.

[Archaeal community structure and diversity in Urumqi No. 10 cold sulfur spring analyzed by culture-independent approach].

OBJECTIVE: We explored the composition and diversity of archaea in a cold sulfur spring water in Xinjiang earthquake fault zone. METHODS: Environmental total DNA was extracted directly with enzymatic lysis method from a cold sulfur spring water. We constructed clone library of 16S rRNA gene amplified with archaeal-specific primers. A total of 115 positive clones were selected randomly from the library and identified by restriction length polymorphism (RFLP) with enzyme Alu I and Afa I. The unique RFLP patterns corresponded clones were selected for sequencing, BLAS alignment and constructing 16S rRNA gene phylogenetic tree. RESULTS: In total, 44 operational taxonomic units (OTUs) were determined from the library. BLAST and phylogenetic analysis indicated that these OTUs were affiliated with Euryarchaeota (94.78%) and Thaumarchaeota (4.35%). Only one Thaumarchaeotal clone was detected and most related to the genus Nitrosopumilus with 93% similarity. Euryarchaeotal clones were abundant and diverse. Of them, 42.61% of clones belonged to RC-V cluster; 13.91% of clones, 20.87% of clones were classified into LDS cluster and Methanomicrobiales respectively; 4.35% of clones had high similarity with ANME-1a-FW, which were involved in Anaerobic oxidation of methane (AOM). In addition, we also detected some (13.05%) unknown Euryarchaotal clones. CONCLUSION: Euryarchaeota in the environment were diverse, and possibly with a large fraction of potential novel species.

Glycyrrhetinic acid extracted from Glycyrrhiza uralensis Fisch. induces the expression of Toll-like receptor 4 in Ana-1 murine macrophages.

Glycyrrhetinic acid (GA) is an active component of licorice root that has long been used as a herbal medicine for the treatment of peptic ulcer, hepatitis, and pulmonary and skin diseases in Asia and Europe. In this study, we analyzed the effect of GA extracted from Glycyrrhiza uralensis Fisch. on the expression of Toll-like receptors (TLRs) that play key roles in regulating the innate immune response against invading pathogens. Stimulation of Ana-1 murine macrophages with GA induced a significant dose-dependent expression of TLR-4, and its mRNA expression that increased from 3-h post-treatment was approximately fivefold over the level in the mock-treated cells. No endotoxin contamination contributed to the GA-induced TLR-4 expression, because polymyxin B treatment did not alter the upregulated expression of TLR-4 in GA-treated cells. Several molecules, such as myeloid differentiation factor 88, interferon-ß, and interleukin-6, which are involved in the TLR-4 downstream signaling pathway, were upregulated significantly in response to GA stimulation. Our findings demonstrate that GA is able to induce the expression of TLR-4 and activate its downstream signaling pathway.

[Bacterial community structure and diversity in a cold sulfur spring in Xinjiang faulting zone].

OBJECTIVE: In order to investigated composition and diversity of bacterial in a cold sulfur spring in Xinjiang faulting zone. METHODS: Environmental total DNA was directly extracted from the water of the No. 10 cold sulfur spring. The 16S rRNA genes were amplified from the total DNA by PCR with bacteria-specific primers and construction a clone library. Positive clones were randomly selected from the library and identified by restriction fragment length polymorphism (RFLP). The unique RFLP pattern corresponded sequences were sequenced, BLAST and then constructed phylogenetic tree. RESULT: In total, 228 positive clones were screened and grouped into 33 Operational Taxonomic Units (OTUs). The clone coverage C value was 92%. 33 Operational Taxonomic Units were divided into 3 phyla with Blast analysis and RDP classifer: Proteobacteria, Bacteroidetes, and Firmicutes. Proteobacteria (98%) was the absolutely dominant group, of which 20% of the clones were highly related to the known photoautotrophic and chemoautotrophic bacteria ( > 97% sequence similarity). Besides, 64% of the clones showed less than 96% of sequence similarity with sequence deposited in GenBank database, of them 54% sequences were affiliated to genus Legionella spp. CONCLUSION: Bacterial diversity in No. 10 cold sulfur spring was low, but maybe have a diversity of novel species and lineages. In addition, large number of novel species of Legionella were detected in the spring water may suggest the water potentially a source of Legionnaires disease and may constitute a menace to the health of human and livestock that lived down the spring.

Icariin induces the expression of toll-like receptor 9 in ana-1 murine macrophages.

Icariin is the major pharmacologically active compound of Herba epimedii which has been used as a tonic, aphrodisiac and an antirheumatic in traditional Chinese medicine. This study analysed the effect of icariin on the expression of Toll-like receptor 9 (TLR9) which plays an important role in regulation of the innate immune response. Stimulation of Ana-1 murine macrophages with icariin induced a significant dose-dependent expression of TLR9, and its mRNA expression which increased from 3 h post-treatment was approximately five-fold that of DMSO-treated cells. Several molecules, such as myeloid differentiation factor 88, tumor necrosis factor-α and interleukin 6, which are involved in the TLR9 downstream signaling pathway, were also significantly up-regulated in response to icariin stimulation. Our findings demonstrated that icariin is able to induce the expression of TLR9.

Design and synthesis of benzoazepin-2-one analogs as allosteric binders targeting the PIF pocket of PDK1.

A novel series of benzoazepin-2-ones were designed and synthesized targeting the PIF pocket of AGC protein kinases, among which a series of thioether-linked benzoazepin-2-ones were discovered to bind to the PIF pocket of 3-phosphoinositide-dependent kinase-1 (PDK1), and to displace the PIF peptide with an EC(50) values in the lower micromolar range. The structure-activity relationships (SARs) of the linker region, tail region, and distal region were explored to further optimize these novel binders which target the PIF pocket of PDK1. When tested in an in vitro PDK1 enzymatic assay using a peptide substrate, the benzodiazepin-2-ones increased the activity of the enzyme in a concentration-dependent fashion, indicating these compounds act as PDK1 allosteric activators. These new compounds may be further developed as therapeutic agents for the treatment of diseases where the PDK1-mediated AGC protein kinases are dysregulated.

An intracellular conformational sensor assay for Abl T315I.

Conformational change is a common molecular mechanism for the regulation of kinase activities. Small molecule modulators of protein conformations, including allosteric kinase inhibitors, are highly wanted as tools for the interrogation of kinase biology and as selective therapeutic agents. However, straightforward cellular assays monitoring kinase conformations in a manner conducive to high-throughput screening (HTS) are not readily available. Here we describe such an HTS-compatible conformational sensor assay for Abl based on a split luciferase construct. The Abl sensor responds to intramolecular structural rearrangements associated with intracellular Abl deactivation and small molecule inhibition. The intact regulatory CAP-SH3-SH2 domain is required for the full functionality of the sensor. Moreover, a T334I Abl mutant (T315I in Abl1a) was found to be particularly well suited for HTS purposes and mechanistic intracellular studies of T334I mutant inhibitors. We expect that the split luciferase-based conformational sensor approach might be more broadly useful to probe the intracellular activation of other kinases and enzymes in general.