Eco-friendly and efficient extraction of polyphenols from Ligustrum robustum by deep eutectic solvent assisted ultrasound.

An eco-friendly and efficient extraction method using deep eutectic solvents assisted ultrasound extraction (DESs-UAE) for the polyphenols from Ligustrum robustum was developed. Among the 34 kinds of DESs prepared, tetraethyl ammonium bromide: 1,2,4-butanol (Teab: 1,2,4-But) was proved to be a suitable extraction solvent based on the extraction efficiency. The extraction parameters including temperature, water content, liquid-solid ratio were optimized with response surface methodology (RSM). Under the optimal conditions, the total phenolic content (TPC) and total flavonoid content (TFC) were 101.46 ± 2.96 mg GAE/g DW and 264.17 ± 5.39 mg RE/g DW, respectively. Furthermore, the extraction mechanism of DESs-UAE was investigated by extraction kinetics, molecular dynamic simulation and theory calculations of interaction. In particular, 9 kinds of polyphenols compounds from Ligustrum robustum were firstly identified by UPLC-Q-TOF-MS. Moreover, the recovered polyphenols exhibited significant antioxidant, α-glucosidase inhibition, acetylcholinesterase inhibition and anticancer activity.

Advances in the Anti-Tumor Activity of Biflavonoids in Selaginella.

Despite the many strategies employed to slow the spread of cancer, the development of new anti-tumor drugs and the minimization of side effects have been major research hotspots in the anti-tumor field. Natural drugs are a huge treasure trove of drug development, and they have been widely used in the clinic as anti-tumor drugs. Selaginella species in the family Selaginellaceae are widely distributed worldwide, and they have been well-documented in clinical practice for the prevention and treatment of cancer. Biflavonoids are the main active ingredients in Selaginella, and they have good biological and anti-tumor activities, which warrant extensive research. The promise of biflavonoids from Selaginella (SFB) in the field of cancer therapy is being realized thanks to new research that offers insights into the multi-targeting therapeutic mechanisms and key signaling pathways. The pharmacological effects of SFB against various cancers in vitro and in vivo are reviewed in this review. In addition, the types and characteristics of biflavonoid structures are described in detail; we also provide a brief summary of the efforts to develop drug delivery systems or combinations to enhance the bioavailability of SFB monomers. In conclusion, SFB species have great potential to be developed as adjuvant or even primary therapeutic agents for cancer, with promising applications.

Lactate Dehydrogenase-Inhibitors Isolated from Ethyl Acetate Extract of Selaginella doederleinii by Using a Rapid Screening Method with Enzyme-Immobilized Magnetic Nanoparticles.

BACKGROUND: Lactate dehydrogenase (LDH) is one of the important enzyme systems for glycolysis and gluconeogenesis. It can catalyze the reduction and oxidation reaction between propionic acid and L-lactic acid, which is usually overexpressed in cancer cells. Therefore, inhibiting the activity of LDH is a promising way for the treatment of cancer. In this study, an effective method based on ligand fishing and ultra performance liquid chromatography-mass spectrum (UPLC-MS) was established to screen and identify active ingredients from Selaginella doederleinii with potential inhibitory activity for LDH. METHODS: Firstly, LDH was immobilized on the magnetic nanoparticles (MNPs), three immobilization parameters including LDH concentration, immobilization time and pH were optimized by single factor and response surface methodology for maximum (max) immobilization yield. Then, a mixed model of galloflavin and chlorogenic acid (inhibitors and non-inhibitors of LDH) was used to verify the specificity of immobilized LDH ligand fishing, and the conditions of ligand fishing were further optimized. Finally, combined with UPLC-MS, immobilized LDH was used to simultaneously screen and identify potential LDH inhibitors from the ethyl acetate extract of Selaginella doederleinii. RESULTS: The prepared fishing material was comprehensively characterized by scanning electron microscopy (SEM), transmission electron microscope (TEM), X-ray diffraction (XRD) and fourier transform infrared spectrometer (FT-IR). The optimal immobilization conditions were obtained as LDH concentration of 0.7 mg/mL, pH value of 4.5, and immobilization time of 3.5 h. Under these conditions, the max immobilization yield was (3.79 ± 0.08) × 103 U/g. The specificity analysis showed that immobilized LDH could recognize and capture ligands, and the optimal ligand fishing conditions included that the incubation time was 30 min, the elution time was 20 min, and the concentration of methanol as eluent was 80%. Finally, two LDH inhibitors, amentoflavone and robustaflavone, were screened by immobilized LDH from the ethyl acetate extract of Selaginella doederleinii. CONCLUSIONS: The study provided a meaningful evidence for discovering the bioactive constituents in ethyl acetate extract of Selaginella doederleinii related to cancer treatment, and this ligand fishing method was feasible for screening enzyme inhibitors from similar complex mixtures.

Purification of biflavonoids from Selaginelladoe derleinii Hieron by special covalent organic polymers material.

Herein, a novel covalent organic polymers (COP) material based on acylhydrazone bond (AB-COP) was prepared as an efficient extraction material for enriching natural medicine biflavonoids from Selaginella doederleinii Hieron. The obtained AB-COP structure was characterized in detail. And it was the first time to investigate the effect of AB-COP on the adsorption of biflavonoids. The effects of initial concentration of solution, adsorption temperature, solid-liquid ratio, adsorption time on the adsorption of biflavonoids were studied. In addition, adsorption kinetic model, adsorption thermodynamic model and density functional theory (DFT) were also investigated to evaluate the adsorption mechanism. At the same time, the static desorption and reusability of AB-COP were investigated. Finally, the dynamic enrichment effect of AB-COP for biflavonoids was investigated. The results showed that AB-COP was successfully synthesized by Fourier transform infrared spectroscopy (FT-IR), solid state nuclear magnetism (NMR), X-ray diffraction (XRD), thermogravimetric analysis (TG), scanning electron microscopy (SEM), laser particle size analysis and Brunner Emmet Teller (BET) specific surface area test. The optimized adsorption parameters of AB-COP were initial concentration of 0.5 mg/mL, temperature of 45 °C, solid-liquid ratio of 10:10 (mg/mL), adsorption time of 60 min. The Langmuir adsorption isotherm could effectively describe the adsorption process, the pseudo-secondary adsorption model could accurately explain the adsorption mechanism, and the DFT calculations revealed that the interaction forces of AB-COP and biflavonoids were π-π stacking and hydrogen bonding. In addition, AB-COP successfully resolved biflavonoids through urea-methanol (1.3 mol/L), and the material can be reused at least four times. Finally, the solid phase extraction (SPE) chromatographic column prepared by AB-COP was successfully applied to the enrichment of biflavonoids from S. doederleinii, and the effect was significantly better than traditional chromatography materials, andthis method was also successfully applied to the enrichment of flavonoids in other plant extracts including Flos sophorae, Pericarpium viride, Lophatheri herba, Herba cuscutae. These results provide references for further purification of bioactive ingredients from plant extracts by using AB-COP.

The Blood Lead Levels of Children and the Loss of Ca2+ from Neurons Owing to Lead.

In order to understand current blood lead levels (BLLs), we investigated the BLLs of children in Sichuan Province from 2011 to 2020. We then monitored the treatment effects of calcium in children with high BLLs to assess their treatment status. Finally, we explored the effects of lead on Ca2+ through in-situ experiments. Whole blood samples were used for BLL tests. The BLLs of 76,362 children aged 0-7 years were measured using atomic absorption spectrometry. The median BLL was 35 µg/L (interquartile range: 28-47). The BLLs were significantly higher in boys than in girls (p < 0.001). The BLLs generally decreased annually and increased with age. The overall prevalence of BLLs ≥ 100 µg/L was 1.20%. The children with high BLLs received subsequent check-ups, and the median time required for effective treatment was 18 months. We observed that lead exposure led to a gradual and persistent loss of Ca2+ levels in neurons of mice brain slices, and the effect did not subside immediately even after the lead was removed. China has made rapid progress in pediatric healthcare, but the treatment status remains unsatisfactory. Because lead causes an irreversible loss of Ca2+, there is an urgent need to develop new standardized treatments to reduce the treatment duration.

Hemolytic Disease of the Fetus and Newborn Caused by Maternal Autoantibody with Mimicking Anti-E Specificity.

OBJECTIVE: There are few reports of hemolytic disease of the fetus and newborn (HDFN) caused by maternal autoantibodies. METHODS: We describe the case of a pregnant patient aged 26 years with systemic lupus erythematosus without any transfusion history who developed autoantibody with mimicking anti-E specificity. Her newborn developed HDFN caused by the maternal autoantibody. RESULTS: The clinical symptoms of the newborn were not serious. After bilirubin light phototherapy and other symptomatic supportive treatment, the baby was discharged with a good prognosis. CONCLUSION: This is the first reported case of HDFN caused by maternal autoantibody with mimicking anti-E specificity. However, the real antigenic target of the autoantibody was not clear.

Effect of probiotic supplementation in pregnant women: a meta-analysis of randomised controlled trials.

Probiotics are being used increasingly in pregnant women, whereas the efficiency on pregnancy outcomes is yet lacking. PubMed, Embase and the Cochrane Library were searched. Relative risks (RR) or weighted mean differences (WMD) with 95 % CI were employed to calculate the summary outcomes. A total of eighteen randomised controlled trials (RCT) including 4356 pregnant women were eligible. The summary RR indicated that probiotic supplementation was associated with a significant decrease in the risk of atopic eczema (RR 0·68; 95 % CI 0·58, 0·81; P < 0·001) and eczema (RR 0·79; 95 % CI 0·68, 0·91; P = 0·002) without significant heterogeneity. Probiotic supplementation was associated with a prolonged gestational age (WMD 0·09; 95 % CI 0·04, 0·15; P = 0·001) with insignificant heterogeneity, whereas no significant effect was exerted on birth weight (P = 0·851). The risks of death (RR 0·34; 95 % CI 0·13, 0·91; P = 0·031) and necrotising enterocolitis (NEC) (RR 0·38; 95 % CI 0·18, 0·81; P = 0·012) were significantly reduced in pregnant women receiving probiotics without evidence of heterogeneity. These findings suggested that probiotics in pregnant women were beneficial for atopic eczema, eczema, gestational age, death and NEC.

Ultrasonic-Assisted Ionic Liquid Extraction of Two Biflavonoids from Selaginella tamariscina.

Selaginella tamariscina, a traditional Chinese medicine, contains a variety of bioactive components, among which biflavonoids are the main active ingredients and have antioxidant, antitumor, and anti-inflammatory properties. In this study, ultrasonic-assisted ionic liquid extraction (UAILE) is used for the first time to extract two main biflavonoids (amentoflavone (AME) and hinokiflavone (HIN)) from S. tamariscina. A high-performance liquid chromatography method is used for the simultaneous determination of AME and HIN in S. tamariscina. Then, three novel ILs are synthesized for the first time by a one-step method using benzoxazole and three acids or acid salts as raw materials, and the structures of the synthesized ILs are characterized by elemental analysis, infrared spectroscopy, and NMR spectroscopy, as well as the thermal stability of the ILs is evaluated by thermogravimetric analysis. After screening the extraction effects of three benzoxazole ILs, three pyridine ILs, and three imidazole ILs, it is found that [Bpy]BF4 is the best and therefore selected as the extractant. The optimal extraction process is explored in terms of the yields of AME and HIN from S. tamariscina by a single-factor experiments and response surface analysis. Under the optimal level of each influencing factor (IL concentration of 0.15 mol/L, solid-liquid ratio of 1:12 g/mL, ultrasonic power of 280 W, ultrasonic time of 30 min, and three extraction cycles), the extraction rates of AME and HIN from S. tamariscina are 13.51 and 6.74 mg/g, respectively. Moreover, the recovery experiment of [Bpy]BF4 on the extraction of biflavonoids shows that the recovered IL can repeatedly extract targets six times and the extraction rate is about 90%, which indicates that the IL can be effectively reused. UAILE can effectively and selectively extract AME and HIN, laying the foundation for the application of S. tamariscina.

Ionic Liquid-Microwave-Based Extraction of Biflavonoids from Selaginella sinensis.

Selaginella sinensis (Desv.) Spring has been used for many years as traditional Chinese medicine (TCM) for many years. Recently, ionic liquids (ILs) have attracted great attentions in extraction and separation technology of TCM as a new green solvent. In this paper, microwave assisted extraction-IL (MAE-IL) that extracted amentoflavone (AME) and hinokiflavone (HIN) from Selaginella sinensis was reported for the first time. The contents of two biflavonoids were simultaneously determined by a high performance liquid chromatography (HPLC) method. After different ionic liquids were compared, it was found [C6mim]BF4 had a high selectivity and efficiency. Moreover, the important extraction conditions, including solid-liquid ratio, IL concentration, extraction time, microwave power and radiation temperature, were also investigated and optimized by response surface methodology (RSM) using AME and HIN yields as index. The results showed that the extraction yields of AME and HIN from S. sinensis were 1.96 mg/g and 0.79 mg/g, respectively, under the optimal process parameters (0.55 mmol/L, 300 W, 40 min, 1:11 g/mL and 48 °C). Compared with the conventional extraction methods, MAE-IL could not only achieve higher yield in shorter time, but also could reduce the consumption of solvent. This effective, rapid and green MAE-IL method was suitable for the extraction of AME and HIN.

Efficacy and safety of FOLFIRI and biotherapy versus FOLFIRI alone for metastatic colorectal cancer patients: A meta-analysis.

BACKGROUND: Previous randomized controlled trials (RCTs) and meta-analyses have demonstrated the useless of FOLFIRI alone for previously treated patients with metastatic colorectal cancer (mCRC). The role of FOLFIRI regimen combined with biological therapy is unknown. The purpose of this meta-analysis is to evaluate the efficacy and safety of combining biological therapy with chemotherapy in previously treated patients with mCRC. METHODS: MEDLINE, EMBASE, Web of Science, Cochrane library, and ClinicalTrials.gov were searched. Eligible studies were RCTs that evaluated the efficacy and safety of the FOLFIRI regimen with or without biological therapy for previously treated patients with mCRC. The hazard ratio (HR) or risk ratio (RR) with 95% confidence interval was estimated. The Chi-squared and I-squared tests were used to assess the statistical heterogeneity. RESULTS: The literature search identified 7 RCTs that met the inclusion criteria for the meta-analysis, and 3680 patients with mCRC were included. The meta-analysis showed that combined therapy was associated with a significant improved progression-free survival (PFS) (HR = 0.78, 95% CI = 0.72-0.85, P < .001), overall survival (OS) (HR = 0.84, 95% CI = 0.77-0.92, P < .001), and overall response rate (ORR) (RR = 1.70, 95% CI = 1.25-2.31, P = .001). Sensitivity analysis suggested that combined therapy versus FOLFIRI alone might increase the risk of Grade 3/4 AEs. CONCLUSION: The addition of biological therapy to the FOLFIRI regimen improved the PFS, OS, and ORR compared with FOLFIRI alone for previously treated patients with mCRC. Long-term survival outcomes are warranted.

The Clinical Characteristics of Neonatal Sepsis Infection in Southwest China.

OBJECTIVE: To identify the pathogens responsible for neonatal sepsis in a high-volume women and children's hospital in Southwest China. METHODS: We retrospectively studied 133 neonates who were admitted to the West China Women and Children's Hospital between 2008 and 2012 for sepsis. The clinical characteristics of the patients were recorded, and the antibiotic sensitivities of the isolated bacteria were determined. RESULTS: All of the included patients had clinical symptoms of sepsis, and subsequent blood cultures confirmed the infection. Almost 80% of patients were infected with coagulase-negative staphylococci (52.8%), Escherichia coli (23.6%), Klebsiella pneumoniae (16.0%) or Staphylococcus aureus (7.5%). Neonates who were infected with gram-negative bacteria, particularly K. pneumoniae, had lower birth weights and were admitted to hospital within 24 hours of birth. Additionally, 87.5% of the isolated K. pneumoniae strains were resistant to third generation cephalosporins. CONCLUSION: Coagulase-negative staphylococci were the most common pathogens found in neonatal sepsis. Moreover, neonatal sepsis caused by gram-negative bacteria was more often observed in newborns of low birth weight. The isolated strains of gram-negative bacteria were highly resistant to cephalosporins. This observation highlights the issue of antibiotic-resistant pathogens in the clinical setting, which poses an added risk to infants presenting with sepsis.

Association of blood lead with calcium, iron, zinc and hemoglobin in children aged 0-7 years: a large population-based study.

Lead exposure in children has received increasing attention from scientists and public health institutions worldwide. Deficiencies of nutritional essential metals can increase the hazard from lead exposure by enhancing absorption and toxicity of dietary lead. Lead, as a ubiquitous toxicity metal, may interact metabolically with nutritional essential metals. A large population-based study was conducted to investigate blood lead, calcium, iron, zinc and hemoglobin levels in healthy children aged 0-7 years. Based on the records, 158 (3.57 %) of 4,429 children had a blood lead levels (BLLs) ≥ 10 µg/dl, and 1,324 (30.30 %) children had a BLL ≥ 7 µg/dl. BLLs in children aged less than 3 years was lower than those in older children. BLLs had a negative correlation with blood calcium and iron level (r = -0.357 and r = -0.070, P < 0.01). Multiple logistic regression analysis also showed that BLLs had a negative correlation with blood calcium and iron. BLLs can be influenced by the status of some essential trace metals in children. Supplement of nutritional elements may help reduce lead absorption. Children with elevated BLLs (≥ 10 µg/dl) were controlled well in recent years in Chengdu. But the burden of reducing BLLs remains extremely arduous, which requires the joint efforts of both government agencies and the public.

Blood lead levels during different trimesters of pregnancy and the possible influencing factors in Chengdu, China.

Lead is a toxic element. It can damage multiple human organs and systems. In present study, we detected the blood lead levels (BLLs) during the whole pregnancy period and 6-12th weeks after delivery and analyzed their influencing factors by healthy pregnant women. We recruited 128 healthy pregnant women absent of pregnancy or obstetric complications or abnormal pregnancy outcomes as the gravida group. The control group consisted of 120 healthy non-pregnant women. The lead concentrations of all the three pregnancy trimesters and postpartum were: 5.95 ± 2.27, 5.51 ± 1.93, 5.57 ± 1.85, and 6.88 ± 1.90 µg/dl; and the mean lead concentration of control group was 6.87 ± 2.29 µg/dl. We found that the BLLs of the gravida group were lower than that of control group during all three trimesters and occupations, supplement nutrition elements, and time of house painted could affect blood lead levels of pregnant women. Lead-related occupations, using cosmetics, and living in a house painted <1 year are risk factors of high BLLs among pregnant women, while calcium, iron, zinc, and milk supplements are protective factors. These may help people especially pregnant women to reduce lead exposure via supplement of calcium, iron, zinc, and milk or avoiding contacting above risk factors.

Environmental lead exposure among children in Chengdu, China, 2007-2009.

Lead poisoning is one of the most significant environmental health threats which children face. The aim of the study was to evaluate the relationship between the blood lead level (BLL) of children and environmental pollution, as well as the effect of BLL on children's heath, in the biggest city in south-west China. A questionnaire survey was administered to 3,624 children's (6 months-7 years old) guardian in urban Chengdu from 2007 to 2009, collecting data on inhabited environment, living habits, and health status. All blood samples were collected for BLL test when the children came to the hospital. The analysis was conducted in West China Second University Hospital, Chengdu, China in December, 2009. A total of 3,624 children were included. The mean BLL was 62.31 µg/L. Three hundred thirty-three (9.2%) of 3,624 children's BLL were above 100 µg/L. Taking Chinese medicinal herbs usually, puffed foods, and substitutes of breast milk were risk factors (P < 0.05).The study shows that Chinese children's lead poisoning prevention and treatment lags far behind developed countries. The findings have implications for environmental health policy.