Cloning and functional validation of DsWRKY6 gene from Desmodium styracifolium.

The purpose of this study was to analyze the role of transcription factor in Desmodium styracifolium, proving that the DsWRKY6 transcription factor was related to the plant phenotypes of Desmodium styracifolium - cv. 'GuangYaoDa1' and it could be used in molecular-assisted breeding. 'GuangYaoDa1' was used as the material and its DNA was the template to clone DsWRKY6, the transgenic Arabidopsis thaliana line was constructed by agrobacterium tumefaciens­mediated transformation. Transgenic Arabidopsis thaliana was cultivated to study phenotype and physiological and biochemical indexes. Phenotypic observation showed that DsWRKY6 transgenic Arabidopsis thaliana had a faster growth rate while compared with the control group, they had longer lengths of main stem, lateral branches of cauline leaves, and root, but a lower number of cauline leaves and lateral branches of cauline leaves. And it also showed that their flowering and fruiting periods were advanced. The results of physiological and biochemical indexes showed that the relative expressions of DsWRKY6 increased and the abscisic acid content significantly increased in DsWRKY6 transgenic Arabidopsis thaliana compared with the control group. According to the above results, DsWRKY6 could regulate the advancing of flowering and fruiting periods caused by the improvement of abscisic acid content, and expression of the DsWRKY6 transcription factor might be the cause of the upright growth of 'GuangYaoDa1'.

Evaluating and optimizing Acid-pH and Direct Lysis RNA extraction for SARS-CoV-2 RNA detection in whole saliva.

COVID-19 has been a global public health and economic challenge. Screening for the SARS-CoV-2 virus has been a key part of disease mitigation while the world continues to move forward, and lessons learned will benefit disease detection beyond COVID-19. Saliva specimen collection offers a less invasive, time- and cost-effective alternative to standard nasopharyngeal swabs. We optimized two different methods of saliva sample processing for RT-qPCR testing. Two methods were optimized to provide two cost-efficient ways to do testing for a minimum of four samples by pooling in a 2.0 mL tube and decrease the need for more highly trained personnel. Acid-pH-based RNA extraction method can be done without the need for expensive kits. Direct Lysis is a quick one-step reaction that can be applied quickly. Our optimized Acid-pH and Direct Lysis protocols are reliable and reproducible, detecting the beta-2 microglobulin (B2M) mRNA in saliva as an internal control from 97 to 96.7% of samples, respectively. The cycle threshold (Ct) values for B2M were significantly higher in the Direct Lysis protocol than in the Acid-pH protocol. The limit of detection for N1 gene was higher in Direct Lysis at ≤ 5 copies/µL than Acid-pH. Saliva samples collected over the course of several days from two COVID-positive individuals demonstrated Ct values for N1 that were consistently higher from Direct Lysis compared to Acid-pH. Collectively, this work supports that each of these techniques can be used to screen for SARS-CoV-2 in saliva for a cost-effective screening platform.

[Hunyuan moxibustion for diarrhea-predominant irritable bowel syndrome of spleen and kidney yang deficiency: a randomized controlled trial].

OBJECTIVE: To compare the therapeutic effect between Hunyuan moxibustion and oral western medication on diarrhea-predominant irritable bowel syndrome（IBS-D）of spleen and kidney yang deficiency. METHODS: Sixty patients with IBS-D of spleen and kidney yang deficiency were randomly divided into a Hunyuan moxibustion group and a western medication group, 30 cases each group. The Hunyuan moxibustion group was treated with Hunyuan moxibustion at Guanyuan（CV 4），40 min each time, once a day; in the western medication group，loperamide hydrochloride capsules (2 mg each time, 3 times a day) and bacillus licheniformis live capsules (0.5 g each time, 3 times a day) were given orally.Both groups were treated for 20 days. The scores of irritable bowel syndrome（IBS）symptom severity scale（IBS-SSS）, IBS quality of life scale (IBS-QOL) and TCM symptom grading quantitative were observed before and after treatment, and the clinical efficacy and safety were evaluated in the two groups. RESULTS: After treatment，each item scores and total scores of IBS-SSS in the two groups were lower than those before treatment（P<0.05）, and the total scores of IBS-QOL were higher than those before treatment (P<0.05)；each item score and total score of IBS-SSS in the Hunyuan moxibustion group were lower than those in the western medication group (P<0.05), and the total score of IBS-QOL in the Hunyuan moxibustion group was higher than that in the western medication group (P<0.05).After treatment, each item score and total score of TCM symptom grading quantitative in the Hunyuan moxibustion group were lower than those before treatment (P<0.05), the abdominal pain, diarrhea, lack of appetite scores and total score in the western medication group were lower than those before treatment (P<0.05)；and the abdominal pain, soreness and weakness of waist and knees, fear to cold and cold limbs scores and total score in the Hunyuan moxibustion group were lower than those in the western medication group (P<0.05).The total effective rate was 90.0%（27/30）in the Hunyuan moxibustion group, which was higher than 73.3%（22/30）in the western medication group (P<0.05). No adverse reactions occurred in both groups during treatment. CONCLUSION: Hunyuan moxibustion can effectively improve the symptom severity and quality of life in patients with IBS-D of spleen and kidney yang deficiency, especially in improving the symptoms of abdominal pain, soreness and weakness of waist and knees, fear to cold and cold limbs.Its therapeutic effect is superior to western medication.

[A correlation study of moxibustion time and moxibustion effect of long snake moxibustion in intervention of recurrent exopathogenic diseases in patients with yang-deficiency constitution].

OBJECTIVE: To explore the time-effect relationship of long snake moxibustion in intervening recurrent exopathogenic diseases of patients with yang-deficiency constitution in different moxibustion periods and provide a scientific basis for the selection of long snake moxibustion in preventing and treating recurrent exopathogenic diseases of patients with yang-deficiency constitution. METHODS: Ninety patients with yang-deficiency constitution who met the inclusion criteria of recurrent exopathogenic diseases were randomly divided into a 30 min group, a 60 min group, and a 90 min group, with 30 cases in each group. Long snake moxibustion was applied once a week from Dazhui (GV14) to Yaoshu (GV2) for different periods (30, 60, and 90 min), 12 times (12 weeks) in total. The scores of yang-deficiency constitution quality scale and Fatigue Severity Scale (FSS) before treatment, after treatment, and six months after treatment, as well as attack times of exopathogenic diseases within one year before treatment and after treatment in the three groups, were observed and recorded. RESULTS: After treatment and 6 months after treatment, the yang-deficiency quality scale scores and FSS scores of the three groups were lower than those before treatment (P<0.01), which decreased in the sequence of the 30 min group, the 90 min group, and the 60 min group (P<0.05). Within one year after treatment, the attack times of exopathogenic diseases in the three groups was lower than that within one year before treatment (P<0.01), which decreased in the sequence of the 30 min group, the 90 min group, and the 60 min group (P<0.05). CONCLUSION: The optimal moxibustion time of long snake moxibustion on the recurrent exopathogenic diseases of patients with yang-deficiency constitution is 60 min.

A one-step molded microfluidic chip featuring a two-layer silver-PDMS microelectrode for dielectrophoretic cell separation.

Dielectrophoresis (DEP) is a powerful technique for label-free cell separation in microfluidics. Easily-fabricated DEP separators with low cost and short turnaround time are in extremely high demand in practical applications, especially clinical usage where disposable devices are needed. DEP separators exploiting microelectrodes made of conducting polydimethylsiloxane (PDMS) composites enable the construction of advantageous 3D volumetric electrodes with a simple soft-lithography process. Yet, existing devices incorporating microelectrodes in conducting PDMS generally have their fluidic sidewalls constructed using a different material, and consequently require extra lithography of a sacrificial layer on the semi-finished master for molding the electrode and fluidic sidewalls in separate steps. Here we demonstrate a novel microfluidic DEP separator with a 3D electrode and fluidic structure entirely integrated within silver-PDMS composites. We develop a further simplified one-step molding process with lower cost using a readily-available and reusable SU8 master, eliminating the need for the additional lithography step in existing techniques. The uniquely designed two-layer electrode exhibits a spatially non-uniform electric field that enables cell migration in the vertical direction. The electrode upper layer then offers a harbor-like region for the trapping of the target cells that have drifted upwards, which shelters them from being dragged away by the main flow streams in the lower layer, and thus allows higher operation flow rate. We also optimize the upper layer thickness as a critical dimension for protecting the trapped cells from high drag and show easy widening of our device by elongation of the digits. We demonstrate that the elongated digits involving more parallel flow paths maintain a high capture efficiency of 95.4% for live cells with 85.6% purity in the separation of live/dead HeLa cells. We also investigate the device feasibility in a viability assay for cells post anti-cancer drug treatment.

[Clinical effect of long-snake moxibustion on lumbago of cold-dampness type].

OBJECTIVE: To observe the effect of long-snake moxibustion on pain, functional disorder and body constitution in the patients with lumbago of cold-dampness type. METHODS: A total of 90 patients with lumbago of cold-dampness type were randomized into long-snake moxibustion, acupuncture and medication groups, 30 cases in each group. In the long-snake moxibustion group, long-snake moxibustion was exerted on the spinal (between Dazhui[GV14] and Yaoshu[GV2]) with the self-prepared moxa powder formula combined with fresh ginger and moxa wool, once a week, for 4 weeks. Regular acupuncture was given to the acupuncture group, 30 min each time, once a day, 8 times as a treatment course, at the interval of 2 days between the courses. The duration of treatment was 1 month. In the medication group, Diclofenac Sodium was prescribed for oral administration, 25 mg each time, three times a day, and Mecobalamine tablets, 0.5 mg each time, 3 times a day. The duration of medication was 1 month. Before and after treatment, the differences were evaluated in the scores of visual analog scale (VAS), the present pain intensity(PPI), Oswestry disability index (ODI) and the scale of yang-deficiency body constitution among the 3 groups. RESULTS: After the treatment, VAS and PPI score and ODI were significantly reduced in the patients of the 3 groups as compared with those before treatment (all P<0.05). Compared with the medication group, ODI score was significantly reduced in the long-snake moxibustion group and the acupuncture group (P<0.05). After the treatment, the scores of the scale of yang-deficiency body constitution were significantly reduced in the long-snake moxibustion group and the acupuncture group (P<0.05) as compared with those before the treatment, but not statistically significant in the medication group in comparison before and after the treatment (P>0.05). The score of the scale of yang-deficiency body constitution in the long-snake moxibustion group was significantly lower than that in the acupuncture group and the medication group (P<0.05). Regarding the clinical effect, the effective rate was 93.33% (28/30) in the long-snake moxibustion group and it was 86.67% (26/30) in the acupuncture group and 73.33% (22/30) in the medication group after the treatment. The effective rate in the long-snake moxibustion group was significantly higher than those in the acupuncture group and the medication group (P<0.05). CONCLUSION: The long-snake moxibustion therapy can achieve significant effect on lumbago of cold-dampness type. This therapy can alleviate pain, relieve the functional disorders and improve the conditions of body constitution in the patients. Hence, it is applicable for the clinical promotion.

Role of coke-bounded environmentally persistent free radicals in phenanthrene degradation by hydrogen peroxide.

Emission of polycyclic aromatic hydrocarbons (PAHs) is accompanied with the discharge of carbonaceous particles during the coke production. To degrade the adsorbed PAHs, hydrogen peroxide (H2O2) was applied as an oxidising agent, which might be activated by the inherent environmentally persistent free radicals (EPFRs) on coke particles. The transformation of phenanthrene (PHE), selected as model molecule, was achieved in H2O2/coke particle system without the addition of additional activating agent. This process consumed the particle-bounded EPFRs, inducing the decreasing of spin density from 1.92 × 1018 to 4.4 × 1017 spins g-1 in 30 min of reaction time. Electron paramagnetic resonance (EPR) technique coupled with spin-trapping agent 5, 5-dimethyl-1-pyrroline-N-oxide (DMPO) was used to probe the potential formation of reactive oxygen species. A higher capture [[Formula: see text]] concentration was observed with larger decreases in EPFRs concentration, indicating that EPFRs were the main contributor to the formation of [Formula: see text]. The obtained results suggested that the activation of H2O2 by EPFRs on coke particles resulted in the generation of hydroxyl radical ([Formula: see text]), which then back-reacted with adsorbed PHE. The finding of this study shed light on a new remediation technology for toxic carbonaceous byproducts discharged during the coke production.

Interaction of Ag+ with soil organic matter: Elucidating the formation of silver nanoparticles.

Naturally silver nanoparticles (AgNPs) have been widely observed in ore deposits, coal, natural water and soil environment. Identifying the source of these naturally AgNPs could be helpful for the elucidation of the geochemical cycle of Ag+ and AgNPs. This paper presents the formation of AgNPs by reducing Ag+ in the presence of soil organic matter (SOM) under various environmentally relevant conditions. The formation of AgNPs associated with various SOM (peat humic acid (PHA), peat fulvic acid (PFA), and commercial humic acids (HA-1 and HA-2)) was determined and compared. The physicochemical properties of the tested SOM were studied by electron paramagnetic resonance (EPR) and attenuated total reflection-infrared (ATR-FTIR) techniques. The formation of AgNPs depended on reductive reactions mediated by SOM. Other influential parameters that influenced the formation of AgNPs included concentrations of Ag+ and SOM and the reaction temperature on AgNPs. The produced AgNPs were characterized by transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS). The mean hydrodynamic diameters of AgNPs associated with PHA and PFA were in range from 2.5 to 15 nm, which were smaller than that produced from HA-1 and HA-2 in the range from 20 to 120 nm. Two different Ag states, i.e., Ag2O and Ag0 species, were observed by XPS technique. The results indicated that the formation of AgNPs depends largely on the types and the properties of natural organic matter. These findings have important implications for the fate of AgNPs under the soil environment.

Sanguinarine inhibits epithelial ovarian cancer development via regulating long non-coding RNA CASC2-EIF4A3 axis and/or inhibiting NF-κB signaling or PI3K/AKT/mTOR pathway.

OBJECTIVE: This study aimed to investigate the antitumor effects and possible regulatory mechanisms of sanguinarine in epithelial ovarian cancer. MATERIAL AND METHODS: The effects of sanguinarine on the malignant behaviors of epithelial ovarian cancer SKOV3 cells and the expression of long non-coding RNA CASC2 were investigated. The expression of CASC2 and EIF4A3 in epithelial ovarian cancer tissues and cells were detected, and the potential mechanisms of sanguinarine were explored by investigating the interactions between CASC2 and EIF4A3. Furthermore, the regulatory relationship between sanguinarine and nuclear factor-κB (NF-κB) signaling or PI3K/AKT/mTOR pathway was explored. RESULTS: Sanguinarine exhibited antitumor effects in SKOV3 cells by significantly inhibiting cell viability, migration and invasion and promoting cell apoptosis. Moreover, sanguinarine induced CASC2 expression and silencing of CASC2 reversed the effects of sanguinarine in epithelial ovarian cancer cells. CASC2 was significantly lowly expressed in ovarian cancer tissues and cells, while EIF4A3 was highly expressed. EIF4A3 was identified as a CASC2 binding protein. Knockdown of EIF4A3 reversed the effects of sanguinarine plus CASC2 silencing. Besides, sanguinarine markedly inhibited the activation of NF-κB signaling or PI3K/AKT/mTOR pathway, which was reversed by CASC2 silencing. And the effects of sanguinarine plus CASC2 silencing on the activation of these pathways were further reversed after knockdown of EIF4A3 at the same time. CONCLUSIONS: Our findings reveal that sanguinarine exhibits antitumor effects in epithelial ovarian cancer cells possible via regulating CASC2-EIF4A3 axis and/or inhibiting NF-κB signaling or PI3K/AKT/mTOR pathway. Sanguinarine may serve as a potential therapeutic reagent for epithelial ovarian cancer.

The impact of joint responses of devices in an airport security system.

In this article, we consider a model for an airport security system in which the declaration of a threat is based on the joint responses of inspection devices. This is in contrast to the typical system in which each check station independently declares a passenger as having a threat or not having a threat. In our framework the declaration of threat/no-threat is based upon the passenger scores at the check stations he/she goes through. To do this we use concepts from classification theory in the field of multivariate statistics analysis and focus on the main objective of minimizing the expected cost of misclassification. The corresponding correct classification and misclassification probabilities can be obtained by using a simulation-based method. After computing the overall false alarm and false clear probabilities, we compare our joint response system with two other independently operated systems. A model that groups passengers in a manner that minimizes the false alarm probability while maintaining the false clear probability within specifications set by a security authority is considered. We also analyze the staffing needs at each check station for such an inspection scheme. An illustrative example is provided along with sensitivity analysis on key model parameters. A discussion is provided on some implementation issues, on the various assumptions made in the analysis, and on potential drawbacks of the approach.