Structure characterization and antioxidant activity analysis of polysaccharides from Lanzhou Lily.

Lanzhou Lily (Lilium davidii var. unicolor) is a traditional medicinal plant and popular edible vegetable bulb in China. In this study, the polysaccharides of Lanzhou Lily (LLPs) were extracted by polyethylene glycol-based ultrasonic-assisted enzymatic extraction method (PEG-UAEE). The optimum process conditions were obtained by single-factor experiments and response surface methodology (RSM). Then, the preliminarily structure of LLPs was characterized by HPLC, FT-IR, and SEM, and its antioxidant activities were evaluated. The results showed that LLPs yield reached 14.75% under the optimized conditions: E/S ratio 1,400 U/g; pH 5.0, ultrasonic time 30 min; and ultrasonic temperature 50 °C. The LLPs has pyranoid ring, uronic acid, and the characteristic absorption peaks of -OH, C = O, and C-H. The results of scanning electron microscope indicated that the LLPs had irregular distribution, dispersed structure, and many holes. The HPLC analysis showed that the LLPs were heteropolysaccharide containing galactose (6.36%), glucose (76.03%), rhamnose (2.02%), and arabinose (7.09%). Moreover, the LLPs showed obvious antioxidant effect in vitro.

Efficacy of Qizi Yusi Pill on Pregnancy Outcomes in Women of Advanced Reproductive Age: A Multicenter, Randomized, Double-Blind, Placebo-Controlled Trial.

OBJECTIVE: To evalvate efficacy of Qizi Yusi Pills (QYP), a Chinese medicine compound preparation, on in vitro fertilization-embryo transfer (IVF-ET) in women of advanced reproductive age. METHODS: This multicenter, randomized, double-blind, placebo-controlled trial was conducted from June 2018 to October 2019. A total of 124 patients were randomly allocated to either the QYP group or the placebo group using a stratified block randomization design, with 62 patients in each group. All patients completed controlled ovarian stimulation using a standard gonadotropin-releasing hormone agonist (GnRH-a) long protocol. As the QYP group, QYP was administered while the control group received placebo. QYP and placebo were administered for a total of 24 to 30 days from the day of GnRH-a pituitary downregulation to transvaginal oocyte retrieval. Both medications were taken orally at doses of 10 g three times each day. The primary outcome was cumulative pregnancy rate, and the secondary outcomes were periodic medication, follicular status, serum hormone and endometrial receptivity. Follow-up continued until 4 weeks after delivery. Maternal and neonatal complications, such as gestational diabetes, were also observed. RESULTS: Overall, 119 patients completed the study, 60 in the QYP group and 59 in the placebo group. Per protocol (PP) analysis revealed that 6-month cumulative pregnancy rate in the QYP group was significantly higher than that in the placebo group [43.33% (26/60) vs. 25.42% (15/59), P=0.040). Additionally, more oocytes were retrieved from the QYP group than those from the placebo group (8.95 ± 3.12 vs. 7.85 ± 1.91, P=0.022). Moreover, the endometrial thickness of HCG day in the QYP group was significantly higher than that in the placebo group (11.78 ± 2.27 mm vs. 10.68 ± 2.07 mm, P=0.012). Maternal and neonatal complications between the two groups were not significantly different (P>0.05). Intention-to-treat analysis was in line with PP results. CONCLUSIONS: QYP can enhance ovarian reserve capacity and ovarian response, and possibly promote endometrial receptivity. QYP effectively improves cumulative pregnancy rates in older patients (⩾35 years) undergoing IVF-ET. (Registration No. ChiCTR1800014427).

The Mechanism of Quercetin in the Treatment of Lung Squamous Cell Carcinoma Based on a Protein-Protein Interaction Network.

Background: Lung squamous cell carcinoma (LUSC) is characterized by poor prognosis and obvious limitations of therapeutic methods. The molecular target and mechanism of quercetin (QR), a natural anticancer product with extensive pharmacological activities, on lung squamous cell carcinoma is still unclear. Method: The effects of QR on LUSC were examined using cell proliferation, migration, and invasion tests. Key target genes were screened using The Cancer Genome Atlas (TCGA) database, Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) database, STRING website, topology, and prognosis analysis, molecular docking, and other bioinformatics methods for further analysis. Finally, the effects of QR on the expression of key targets in LUSC cells were detected using a cell cycle assay and western blotting. Results: Our study demonstrates that QR not only inhibits the proliferation of LUSC but also affects the invasion and metastasis of LUSC. After downloading and analyzing the TCGA database, 2150 differentially expressed genes were identified. PLK1, CDC20, and BUB1B were identified using enrichment analysis, topological network analysis, cluster analysis, and molecular docking screening. Subsequent experiments showed that QR could interfere with the cell cycle and downregulate the expression of the target gene PLK1 at the protein level. Conclusions: We found that QR not only inhibits the proliferation, migration, and invasion but also blocks the cell cycle progression of LUSC. QR downregulated the expression of the LUSC target gene PLK1 at the protein level.

Effect of traditional Chinese medicine formula Guilu Xian on in vitro fertilization and embryo transfer outcome in older women with low prognosis: study protocol for a prospective, multicenter, randomized double-blind study.

INTRODUCTION: In recent years, the prevalence of infertility has significantly increased and has become a global reproductive health problem. The female ovarian reserves have been shown to decrease progressively with an increase in age. Besides, the rate of embryo implantation and clinical pregnancy also decreases. Traditional Chinese medicine has been widely applied in assisted reproductive technology. It is reported to have a significant influence on improving the quality of oocytes, improving endometrial receptivity, increasing clinical pregnancy rate, reducing pregnancy-related complications, etc. Therefore, this study will investigate the effect of Guilu Xian, a traditional Chinese medicine formula on IVF-ET outcome in older women with low prognosis. METHODS AND ANALYSIS: This trial is a prospective, multicenter, randomized double-blind clinical trial. A total of 120 infertile patients with low prognosis and receiving IVF or ICSI in 3 public hospitals in China will be randomly divided into two parallel groups: Guilu Xian group (n = 60) and placebo group (n = 60). Patients in both groups will be treated with antagonist regimens to promote ovulation, and all the patients will be required to take the medication from the 2nd to 4th day of the menstrual cycle to the day of egg retrieval. A comparison of the total number of oocytes obtained, the fertilization rate, clinical pregnancy rate, embryo quality, embryo implantation rate, and early spontaneous abortion rate between the experimental group and the placebo group will be performed. TRIAL REGISTRATION: Chinese Clinical Trials Registry ChiCTR1900028255 . Registered on 16 December 2019.

The Role of Traditional Chinese Formula Ding-Kun Pill (DKP) in Expected Poor Ovarian Response Women (POSEIDON Group 4) Undergoing In Vitro Fertilization-Embryo Transfer: A Multicenter, Randomized, Double-Blind, Placebo-Controlled Trial.

Objective: The primary objective of the study was to assess traditional Chinese formula DKP supplementation in terms of efficacy and safety on reproductive outcomes of expected poor ovarian responder (POR, POSEIDON Group 4) undergoing in vitro fertilization-embryo transfer (IVF-ET). Design Setting and Participants: Women eligible for IVF-ET were invited to participate in this randomized, double-blind, placebo-controlled, superiority trial at academic fertility centers of ten public hospitals in Chinese Mainland. A total of 462 patients (35-44 years) equally divided between DKP and placebo groups with antral follicle count (AFC) <5 or anti-müllerian hormone (AMH) <1.2 ng/ml were randomized. Interventions: All participants were given DKP or 7 g placebo twice daily on the previous menstrual cycle day 5 until oocyte retrieval, which took approximately 5 to 6 weeks. Main Outcome Measure: The primary outcome was the ongoing pregnancy defined as more than 20 gestational weeks of an intrauterine living fetus confirmed by pelvic ultrasonography. Results: Demographic characteristics were equally distributed between the study populations. Intention-to-treat (ITT) analysis revealed that ongoing pregnancy rate (OPR) was not significantly different between DKP and placebo groups [26.4% (61/231) versus 24.2% (56/231); relative risk (RR) 1.09, 95% confidence interval (CI) 0.80 to 1.49, P = 0.593]. No significant differences between groups were observed for the secondary outcomes. The additional per protocol (PP) analysis was in line with ITT results: OPR in DKP group was 27.2% (61/224) versus 24.1% (55/228) in placebo group [RR 1.13, 95%CI (0.82 to 1.55), P = 0.449]. After subgroup analysis the findings concluded that POR population of 35-37 years had a significantly higher OPR after 5-6 weeks of oral DKP (41.8%, 33/79) versus placebo (25.4%, 18/71) [RR 1.65, 95% CI (1.02 to 2.65), P = 0.034, P for interaction = 0.028]. Conclusion: This well-designed randomized controlled trial (RCT) offers new high-quality evidence to supplement existing retrospective literature concerning DKP performance in expected PORs. DKP could be recommended as a safe and natural remedy for expected PORs (aged 35-37 years) who fulfill the POSEIDON group 4 criteria. However, additional interventional clinical studies are undoubtedly required to be conducted in the future to validate this hypothesis. Clinical Trial Registration: www.chictr.org.cn, identifier ChiCTR1900026614.

A chemiluminescence assay for determination of lysozyme based on the use of magnetic alginate-aptamer composition and hemin@HKUST-1.

Lysozyme aptamer-functionalized magnetic alginate hydrogel was prepared for separation and enrichment of lysozyme. Luminol-labeled aptamer was used as a signal tag, and the signal tag was adsorbed on magnetic carboxylated carbon nanotubes based on the π-interaction. When lysozyme was added, the aptamer specifically binds to the lysozyme, causing the signal tag to detach from the magnetic carboxylated carbon nanotubes. When the aptamer/lysozyme complex bound to the complementary single strand of aptamer on the hemin@HKUST-1, lysozyme was released. The released lysozyme can be recombined with the signal tag adsorbed on the magnetic carboxylated carbon nanotube, allowing more signal tag to be dispersed into the solution. Determination of lysozyme was achieved by releasing the luminol-labeled aptamer to generate a chemiluminescence signal at a wavelength of 425 nm. It was proved by experiments that the synthesized hemin@HKUST-1 had a strong catalytic effect on the luminol-NaOH-H2O2 system. The chemiluminescence signal was increased nearly 100 times. The complementary pairing allowed the luminol to be immobilized on the surface of hemin@HKUST-1. The generation and consumption of short-lived reactive oxygen species were concentrated on the surface of the MOFs, which improves the chemiluminescence efficiency. The introduction of hemin@HKUST-1 and DNA solved the defects of chemiluminescence analysis. The chemiluminescence assay was able to detect lysozyme with linear range of 1.05 × 10-6 U∙mg-1 (6.00 × 10-13 mol∙L-1)-1.25 × 10-2 U∙mg-1 (7.14 × 10-9 mol∙L-1); the detection limit was 3.50 × 10-7 U∙mg-1 (2.00 × 10-13 mol∙L-1) (R2 = 0.99). The recovery of lysozyme in spiked saliva samples was 97.4-102.8%. Graphical abstract Schematic presentation of chemiluminescence assay. Lysozyme (Lys) was captured by aptamer-modified magnetic sodium alginate (M-Alg-Apt); Glycine (pH = 2) as eluent for Lys. Luminol-modified Apt (Apt-luminol) as signal tag; magnetic carbon nanotubes (MCNTs) as adsorption matrix; cDNA was complementary to Apt; hemin@HKUST-1 as catalyst.

A "signal-on" chemiluminescence biosensor for thrombin detection based on DNA functionalized magnetic sodium alginate hydrogel and metalloporphyrinic metal-organic framework nanosheets.

A "signal-on" chemiluminescence biosensor was established for detecting thrombin. The thrombin aptamer1-functionalized magnetic sodium alginate (Malg-Apt1) hydrogel was synthesized by physical interaction between sodium alginate and Ca2+, and it was used in the biosensor for separating and enriching thrombin. Ethylenediamine tetraacetic acid (EDTA) was used to chelate with Ca2+ to dissolve the hydrogel and release thrombin. A metalloporphyrinic metal-organic framework nanosheet, named as Cu-TCPP(Co) MOFs, was prepared as signal amplification strategy. Cu-TCPP(Co) MOFs/Au-ssDNA (ssDNA: single-strand DNA) was synthesized for controllable further amplification of chemiluminescent signal. The thrombin aptamer2-functionalized magnetic carbon nanotubes (MCNTs-Apt2) were used as a matrix, and Cu-TCPP(Co) MOFs/Au-ssDNA was adsorbed on the MCNTs by the complementary pairing of the partial bases between ssDNA and Apt2. Compared with ssDNA, Apt2 has a stronger interaction with thrombin. Therefore, thrombin can trigger the release of Cu-TCPP(Co) MOFs/Au-ssDNA to achieve signal amplification. Under the optimal conditions, the biosensor could detect thrombin as low as 2.178 × 10-13 mol/L with the range from 8.934 × 10-13 to 5.956 × 10-10 mol/L and exhibited excellent selectively. Moreover, the "signal-on" chemiluminescence biosensor showed potential application for the detection of thrombin in body fluids.

[Differential Responses of Rhizospheric nirK- and nirS-type Denitrifier Communities to Different Phosphorus Levels in Paddy Soil].

Phosphorus is an essential life element, which can affect the activities and functions of denitrifiers. Both nirK and nirS genes can code nitrite reductase; however, it remains unclear whether nirK- and nirS-containing denitrifers respond differentially to changes in the availability of phosphorus in paddy soil. In this study, P-deficient paddy soil was used to grow rice plants. Three phosphorus levels established by applying P fertilizer at a rate of 0 mg·kg-1 (CK), 15 mg·kg-1 (P1), and 30 mg·kg-1(P2), respectively. The abundance and community structure of nirK- and nirS- containing denitrifers were determined using quantitative PCR and high-throughput sequencing techniques. Results indicated that nirK- and nirS-containing communities responded differentially to changes in the P levels. The nirS-containing communities are more sensitive to the changes in P in both rhizosphere and bulk soil samples. In addition, the abundance of nirS genes was 2-3 times higher in the P2 treatment than in the CK treatment. Furthermore, the nirS community structure is also clearly differed from the CK treatment. However, P addition only induced partial modification of the community structure and abundance of nirK-containing denitrifiers. Moreover, compared to the bulk soil with each phosphorus level, the nirS community structure in the rhizosphere soil changed significantly; however, only the P2 treatment induced significant increases in the abundance of the nirS gene. In contrast, no significant differences in the abundance and composition of nirK-containing denitrifers were detected between rhizosphere and bulk soils under different phosphorus levels. Collectively, application of phosphate fertilizer in P-deficient paddy soil could significantly increase the abundance of nirK- and nirS-containing denitrifiers, changing their community structures, with nirS-type showing a greater sensitivity than nirK-type denitrifiers. In comparison, the denitrifying communities in the rhizosphere were more sensitive to variable P levels than that in the bulk soil. Compared to bulk soils, rice growth shifted the community structure of nirS- and nirK-containing denitrifiers in rhizosphere soils at each level of P, but failed to induce significant changes in their abundance (except for P2) that could cause a significant increase in nirS abundance. These results could provide a theoretical basis for exploring the effects of fertilization on soil denitrification.