Active Components, Antioxidant, Inhibition on Metabolic Syndrome Related Enzymes, and Monthly Variations in Mature Leaf Hawk Tea.

Hawk tea is a rich and edible resource, traditionally used as a beverage in South China. This drink has many pharmacologic effects, such as acting as an antioxidant and reducing blood sugar and lipids. The objective of this work was to explore the active compound contents, bioactivities and their monthly changes, and optimize the harvest time. In the present study, Hawk tea from each month in 2017 was collected and extracted with 70% (v/v) ethanol. The contents of the total flavonoids and total phenols were determined using the colorimetric method. We determined the contents of seven characteristic active substances-hyperin, isoquercitrin, trifolin, quercitrin, astragalin, quercetin, and kaempferol-using high-performance liquid chromatography. The crude extract was tested for its antioxidant and inhibitory properties on enzymes involved in metabolic syndrome. Specifically, 2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid), ferric-reducing power assay, and the inhibition capacity test on α-glucosidase and lipase were conducted to determine the antioxidant effect in vitro, as well as the reduction of blood sugar and lipids. Monthly variations in activities and components were determined by numeric analysis and comparison. Correlation analysis revealed that antioxidant effects are significantly correlated with the total flavonoids. The hierarchical cluster analysis of bioactivities and their contents indicates that October and November are the best harvesting months, which differs with the habitual collection of Hawk tea.

Chemical Components, Biological Activities, and Toxicological Evaluation of the Fruit (Aril) of Two Precious Plant Species from Genus Taxus.

The fruit (aril) of the endangered genus Taxus plants is an abandoned herbal resource. Traditionally, people enthusiastically focus on its bark, its renewable, tremendous arils fall into the soil with seeds after they are mature. The present research investigated the fruit of two species from the genus Taxus, Taxus chinensis var. mairei, and Taxus media, with regards to their antioxidant and antihyperglycaemic activities, safety, and bioactive constituents. Results showed that T. chinensis var. mairei and T. media both had certain biological activities with T. chinensis var. mairei better in antioxidant activity and T. media better in antihyperglycaemic activity. Correlation analysis revealed that the differences in bioactivities depended on content of their mainly chemical components. The mice acute oral toxicity test indicated that the methanol extracts of the two biotypes of Taxus were safe. And nineteen compounds were tentatively assigned from the two varieties, via tandem mass spectrometry using a LC-ESI-Q-TOF-MS instrument, which included phenols, flavonoids, and terpenes. These results indicate a possible application of Taxus fruit extracts in various fields like in food industry, however, this still needs further investigations.

[Identification of dominant microbes from traditional Chinese medicine Pinelliae Rhizoma Fermentata].

To investigate the microbial species, amount changes as well as the isolation and identification of domain strains at different fermentation time points of Pinelliae Rhizoma Fermentata, and provide basis for exploring the mechanism of Pinelliae Rhizoma Fermentata processing. Five samples were chosen at the time points (0, 18, 36, 54, 72 h) of Pinelliae Rhizoma Fermentata processing. Bacteria, mold and yeast from the samples were cultured; their colonies were counted, and the dominant strains were isolated and purified. The dominant bacteria and dominant fungi were identified by 16S rDNA and 26S rDNA sequencing respectively. The results showed that the bacteria count was low with slow and smooth changes in the fermentation process;while mold and yeast grew dramatically after 54 h culturing and reached 1×107 CFU•mL⁻¹ at the end of fermentation. Through the NCBI homology alignment and phylogenetic tree construction, the dominant bacteria were identified as Streptomyces sp., Bacillus pumilus, B. subtilis, B. aryabhattai and other Bacillus sp.; the dominant yeast was identified as Meyerozyma guilliermondii; the dominant mold were identified as Paecilomyces variotii, Byssochlamys spectabilis, and Aspergillus niger in the processing of Pinelliae Rhizoma Fermentata. The results indicated that multiple microbe species, especially yeast and mold, played a role in the fermentation processing of Pinelliae Rhizoma Fermentata. M. guilliermondii, P. variotii, P. variotii and A. niger and Bacillus sp. can be the crucial factors in the processing of Pinelliae Rhizoma Fermentata.

[Effect of protein intervention on amino acid metabolism spectrum of Qi and Yin deficiency type 2 diabetic rats].

To study the effect of plant protein and animal protein on amino acid metabolism spectrum of Qi and Yin deficiency type 2 diabetic rats. 110 male SD rats were randomly divided into blank group (n=10), diabetic model group (n=20), disease-symptoms group (n=80). The rats of blank group received ordinary feeding, while other groups were fed with high sugar and fat diets. During the whole process of feeding, rats of disease-symptoms group were given with Qingpi-Fuzi (15.75 g•kg⁻¹) once a day through oral administration. Five weeks later, the rats were given with a low dose of STZ (40 mg•kg⁻¹) by intraperitoneal injection to establish experimental diabetic models. Then the models were randomly divided into disease-symptoms group 1 (Qi and Yin deficiency diabetic group, 15.75 g•kg⁻¹), disease-symptoms group 2 (plant protein group, 0.5 g•kg⁻¹), disease-symptoms group 3 (animal protein group, 0.5 g•kg⁻¹), disease-symptoms group 4 (berberine group, 0.1 g•kg⁻¹). The drugs were given for 4 weeks by gavage administration. After 4 weeks of protein intervention, the abdominal aortic blood was collected and serum was isolated to analyze its free amino acid by using AQC pre-column derivatization HPLC and fluorescence detector. Four weeks after the protein intervention, plant protein, animal protein and berberine had no obvious effect on body weight and blood sugar in type 2 diabetic rats. As compared with animal protein group, histidine and proline（P<0.01）, serine, glycine, threonine, alanine, tyrosine, valine, methionine, bright+isoleucine, phenylalanine and lysine（P<0.05）changed a lot in rats serum of plant protein group.The results showed that gavage administration of protein would produce effects on amino acid metabolism of Qi and Yin deficiency type 2 diabetic SD rats. Symbolic differential compounds could be found through metabonomics technology, providing experimental basis for early warning of type 2 diabetes and diagnosis of Qi and Yin deficiency syndrome.

[Effects of caffeine citrate on myelin basic protein in neonatal rats with hypoxic-ischemic brain damage].

OBJECTIVE: To study the effects of caffeine citrate on myelin basic protein (MBP) expression in the cerebral white matter of neonatal rats with hypoxic-ischemic brain damage (HIBD) and the related mechanism. METHODS: Forty-eight seven-day-old Sprague-Dawley neonatal rats were randomly assigned to 3 groups: sham operation (n=16), HIBD (n=16) and HIBD+caffeine citrate (n=16). The rats in the HIBD and HIBD+caffeine citrate groups were subjected to left common carotid artery ligation, and then were exposed to 80 mL/L oxygen and 920 mL/L nitrogen for 2 hours to induce HIBD. The rats in the sham operation group were only subjected to a sham operation, without the left common carotid artery ligation or hypoxia exposure. Caffeine citrate (20 mg/kg) was injected intraperitoneally before hypoxia ischemia (HI) and immediately, 24 hours, 48 hours and 72 hours after HI. The other two groups were injected intraperitoneally with an equal volume of normal saline at the corresponding time points. On postnatal day 12, the expression of MBP in the left subcortical white matter was detected by immunohistochemistry, and the levels of adenosine A1 receptor mRNA and A2a receptor mRNA in the left brain were detected by real-time PCR. RESULTS: The expression of MBP in the left subcortical white matter in the HIBD group was lower than in the sham operation group (P<0.05). The MBP expression in the HIBD+caffeine citrate group was significantly higher than in the HIBD group, but was still lower than the sham operation group (P<0.05). Real-time PCR showed that the adenosine A1 receptor mRNA expression was significantly higher in the HIBD group than in the sham operation group, and it was significantly lower in the HIBD+caffeine citrate group than in the HIBD group (P<0.05). CONCLUSIONS: Caffeine citrate can improve brain white matter damage following HIBD in neonatal rats and the protection mechanism might be related with the down-regulation of adenosine A1 receptor expression.

[Effects of erythropoietin on neuronal proliferation and apoptosis in neonatal rats after infection-induced brain injury].

OBJECTIVE: To investigate the effects of erythropoietin (EPO) on the neuronal proliferation and apoptosis in neonatal rats after infection-induced brain injury and the neuroprotective mechanism of EPO in neonatal rats with infection-induced brain injury. METHODS: Twenty-six two-day-old neonatal rats were randomly divided into 3 groups: control group (intraperitoneally given an equal volume of normal saline), lipopolysaccharide (LPS) group (intraperitoneally given LPS 0.6 mg/kg), and EPO group (intraperitoneally given LPS 0.6 mg/kg and EPO 5 000 U/kg). These groups were injected with respective drugs for 5 consecutive days. Meanwhile, each group was intraperitoneally injected with 5-bromo-2'-deoxyuridine (BrdU) (50 mg/kg) once a day for 5 consecutive days. The expression of BrdU and cleaved Caspase-3 in the hippocampal dentate gyrus was detected by immunohistochemistry at 24 hours after the last injection. RESULTS: The number of neuronal cells in the hippocampal dentate gyrus in the LPS and EPO groups was significantly greater than in the control group (P<0.05), but there was no significant difference between the LPS and EPO groups. The EPO group had a significantly higher number of BrdU-positive cells in the subgranular zone of hippocampal dentate gyrus than the LPS group (51±9 vs 29±6; P<0.05), but a significantly lower number of BrdU-positive cells than the control group (51±9 vs 67±12; P<0.05). The EPO group had a significantly lower number of cleaved Caspase-3-positive cells in the subgranular zone of hippocampal dentate gyrus than the LPS group (27.9±1.5 vs 34.0±1.3; P<0.05), but a significantly higher number of cleaved Caspase-3-positive cells than the control group (27.9±1.5 vs 21.0±1.7; P<0.05). CONCLUSIONS: EPO can promote hippocampal neuronal proliferation and reduce neuronal apoptosis in neonatal rats after infection-induced brain injury.

[Two-stage Inhibition Effects of Burkholderia sp. Y4 Application on Cadmium Uptake and Transport in Wheat].

In order to evaluate the feasibility of using Burkholderia sp. Y4 as a cadmium （Cd）-reducing bacterial agent in contaminated wheat fields， the changes in the rhizosphere soil microbial community and Cd available state， as well as the content and transport characteristics of Cd in the wheat root， basal node， internode， and grain under the treatment of strain Y4 were tested using microbial high-throughput sequencing， step-by-step extraction， subcellular distribution， and occurrence analyses. The results showed that root application of strain Y4 significantly reduced the root and grain Cd content of wheat by 7.7% and 30.3%， respectively， compared with that in the control treatment. The Cd content and Cd transfer factor results in wheat vegetative organs showed that strain Y4 reduced the Cd transfer factor from basal node to internode by 79.3%， and Cd content in the wheat internode stem also decreased by 50.9%. The study of Cd occurrence morphology showed that strain Y4 treatment increased the proportion of residual Cd in roots and basal ganglia， decreased the contents of inorganic and water-soluble Cd in roots， and increased the content of residual Cd in basal ganglia. Further examination of the subcellular distribution of Cd showed that the Cd content in root cell walls and basal ganglia cell fluid increased by 21.3% and 98.2%， respectively， indicating that the Cd fixation ability of root cell walls and basal ganglia cell fluid was improved by the strain Y4 treatment. In the rhizosphere soil， it was found that the microbial community structure was changed by strain Y4 application. Under the Y4 treatment， the relative abundance of Burkholderia increased from 9.6% to 11.5%， whereas that of Acidobacteriota decreased. Additionally， the relative abundance of Gemmatimonadales， Pseudomonadales， and Chitinophagales were also increased by strain Y4 treatment. At the same time， the application of strain Y4 increased the pH value of rhizosphere soil by 8.3%. The contents of exchangeable Cd， carbonate-bound Cd， and iron-manganese oxide-bound Cd in the soil decreased by 44.4%， 21.7%， and 15.9%， respectively， whereas the proportion of residual Cd reached 53.6%. Root application of strain Y4 increased the contents of nitrate nitrogen and ammonium nitrogen in the soil by 22.0% and 21.4%， respectively， and the contents of alkaline nitrogen also increased to a certain extent. In conclusion， the root application of strain Y4 not only improved soil nitrogen availability but also inhibited Cd transport and accumulation from contaminated soil to wheat grains in a "two-stage" manner by reducing Cd availability in rhizosphere soil and improving Cd interception and fixation capacity of wheat roots and basal nodes. Therefore， Burkholderia Y4 has application potential as a Cd-reducing and growth-promoting agent in wheat.

[Pollution Characteristics and Risk Assessment of PAHs in the Soil of Wild Forsythia Suspensa in Shanxi].

Shanxi is one of the main producing areas of Forsythia suspensa in China. In order to explore the safety of the soil in the areas where Forsythia suspensa grows,70 surface (0-25 cm) soil samples were collected from the main growing areas of F. suspensa in the eastsouth of Shanxi Province in July 2017. The concentration and composition characteristics of 16 polycyclic aromatic hydrocarbons (PAHs) in the sample soils were analyzed using chemical extraction and gas chromatography-mass spectrometry (GC-MS). The diagnostic ratio method was used to determine the source of PAHs in the areas. The potential ecological risk was assessed by using the method of calculating the equivalent carcinogenic concentration of benzo[a]pyrene. The results showed that the average concentration of total PAHs (Σ16PAHs) in all of the soil samples was 1.85 µg·g-1, which was dominated by three ring number PAHs, accounting for 76.7% of the total PAHs. The detection rates of phenanthrene (Phe) and anthracene (Ant) were both 100% of all the sample sites. The soil PAHs in the wild F. suspensa growing areas mainly originated from coal, biomass burning, and motor vehicle exhaust emissions, which resulted from air transport and sedimentation pathways. In all of the sample sites, the concentration of Σ16PAHs the limit standard level (0.2 µg·g-1) of Maliszewska-Kordybach for agricultural soil pollution and exceeded the soil heavy pollution level limit value (1.0 µg·g-1) in 41.4% of the sample sites. The concentration of BaP was above the risk control standard for soil contamination of agricultural land (0.55 µg·g-1) in 10% of all the soil samples. A total of 11.4% of the sample soil ΣBaPeq16PAHs and ΣBaPeq8BPAHs exceeded the agricultural soil screening value (0.55 µg·g-1). These results indicate that the contamination of PAHs was at a detectable level in the soil of wild F. suspensa growing in Shanxi, and thus their potential ecological risks should not be ignored. It is necessary to enhance the research regarding these areas to ensure the safe production of medicinal plants.

[Risk factors for periventricular-intraventricular hemorrhage in premature infants treated with mechanical ventilation].

OBJECTIVE: To study risk factors for periventricular-intraventricular hemorrhage (PVH-IVH) in premature infants treated with mechanical ventilation. METHODS: A total of 205 premature infants who were admitted to the neonatal intensive care unit (NICU) and treated with mechanical ventilation between January 2009 and December 2011 were enrolled. They were classified into PVH-IVH and non-PVH-IVH groups according to the results of head ultrasonography performed at 3 to 7 days after birth. Single factor and multivariate logistic regression analysis were used to identify risk factors for PVH-IVH. RESULTS: Single factor analysis indicated 9 factors associated with the development of PVH-IVH, including a gestational age of <32 weeks, a birth weight of <1500 g, intrauterine distress, severe asphyxia, vaginal delivery, maternal perinatal infection, premature rupture of membranes (PROM) at ≥8 hours, mechanical ventilation duration of ≥7 days and ventilator-associated pneumonia (VAP) (P<0.05). Multivariate logistic regression analysis showed that a birth weight of <1500 g (OR=2.665), intrauterine distress (OR=2.177), severe asphyxia (OR=5.653), maternal perinatal infection (OR=4.365) and VAP (OR=2.299) were independent risk factors for the development of PVH-IVH (P<0.05). CONCLUSIONS: Very low birth weight, intrauterine distress, severe asphyxia, maternal perinatal infection and VAP are closely associated with an increased risk of PVH-IVH in premature infants treated with mechanical ventilation. These clinical risk factors should be given more attention in the prevention of PVH-IVH.

Dose-Response Effects of Low-Dose Ionizing Radiation on Blood Parameters in Industrial Irradiation Workers.

While previous studies have focused on the health effects of occupational exposure of radiations on medical radiation workers, few have analyzed the dose-response relationship between low radiation doses and changes in blood parameters. Even fewer studies have been conducted on industrial worker populations. Using a prospective cohort study design, this study collected health examination reports and personal dose monitoring data from 705 industrial irradiation workers who underwent regular physical examinations at Dongguan Sixth People's Hospital. The dose-response effects of low-dose ionizing radiation on blood parameters were assessed using a generalized linear model and restricted cubic spline model. Red blood cell counts decreased then increased, before decreasing again with increasing ionizing radiation. This was in contrast to the curve of the total platelet count after irradiation. Additionally, a radiation dose of 2.904 mSv was the turning point for the nonlinear curve of hemoglobin count changes. In conclusion, long-term, low-dose ionizing radiation affects blood cell levels in industrial irradiation workers. There is a nonlinear dose-response relationship between red blood cell, platelet, and hemoglobin counts and the cumulative radiation dose. These findings should alert radiation workers to seek preventive medical treatment before the occurrence of any serious hematopoietic disease.

[Evaluation of the Potential Agricultural Risks of Polycyclic Aromatic Hydrocarbon Contaminated Soil by Planting Lactuca sativa L.]

In order to evaluate the potential agricultural risks of soil contaminated by polycyclic aromatic hydrocarbons (PAHs), Lactuca sativa L. was used as a model leaf vegetable plant to investigate the enrichment characteristics of PAHs in the different tissues of Lactuca sativa L, such as its underground parts (GS) and aboveground parts (YS), which were studied through an experiment involving potted cultivation in PAHs contaminated soil that was taken from the agricultural soil around a coking enterprise area. The concentrations of the different PAHs in the soil and plant tissues were analyzed using ultrasonic oscillation extraction and high-performance liquid chromatography-mass spectrometry (GC-MS) analysis methods. The results show that the enrichment of the total PAHs (Σ16PAHs) in the YS is higher than that in the GS. The components enriched in the YS mainly consist of 3-5 ring PAHs, and those in the GS consist of 4-6ring PAHs. The coefficients of the different PAHs enriched in the YS were higher than those of the GS. The enrichment coefficient of anthracene (Ant) was the largest and that of fluoranthene (Fla) was the smallest in the YS, while the enrichment coefficient of benzene[a]pyre (Bap) was the largest and that of Fla was the smallest in the GS. The transfer coefficients of the different PAHs from the GS to the YS were greater than the rate from the initial soil (SS) to the GS; the value is less than 1 from the SS to GS. The correlations and goodness of fit were analyzed for the concentrations of PAHs in the SS, GS and YS. The Σ16PAHs in the SS showed positive correlations with the Σ16PAHs in the YS and in GS. The goodness of fit of the correlation of the PAHs in YS to those in the SS was the largest (R2=1.0), while the goodness-of-fit for the correlation between the GS and SS was lower (R2=0.71). The benzo[a]pyrene equivalent concentration values (Σ16PAHsBapeq) of YS and GS were 11.8 and 12.7 times the maximum value of Bap level allowed in food. The study indicates that contaminated farmland soil would present a high health risk when it was used to grow the plant.

Transfer learning on T1-weighted images for brain age estimation.

Due to both the hidden nature and the irreversibility of Alzheimers disease (AD), it has become the killer of the elderly and is thus the focus of much attention in the medical field. Radiologists compare the predicted brain age with the ground truth in order to provide a preliminary analysis of AD, which helps doctors diagnose AD as early in its development as possible. In this paper, a transfer learning-based method of predicting brain age using MR images and dataset of a public brain was proposed. In order to get the best transfer results, we froze different layers and only fine-tuned the remaining layers. We used three planes of brain MR images together to predict age for the first time and experiment results showed that the proposed method performs better than the state-of-the-art method under mean absolute error metric by 0.6 years. In addition, to explore the relationship between brain MR images of different planes and predicted age accuracy, we used three different planes of brain MR images to predict age respectively for the first time and found that sagittal plane MR images outperformed two other planes in age estimation. Finally, our research identified, the effective regions that contribute to brain age estimation for cognitively normal individuals and for AD patients with deep learning. For AD patients, the effective region is mainly concentrated in the frontal lobe of the brain, verifying the relevant medical conclusions about AD.

Fungal communities in sediments of subtropical Chinese seas as estimated by DNA metabarcoding.

Ribosomal RNA internal transcribed spacer-1 (ITS1) metabarcoding was used to investigate the distribution patterns of fungal communities and the factors influencing these patterns in subtropical Chinese seas, including the southern and northern Yellow Sea and the Bohai Sea. These seas were found to harbor high levels of fungal diversity, with 816 operational taxonomic units (OTUs) that span 130 known genera, 36 orders, 14 classes and 5 phyla. Ascomycota was the most abundant phylum, containing 72.18% and 79.61% of all OTUs and sequences, respectively, followed by Basidiomycota (19.98%, 18.64%), Zygomycota (1.10%, 0.11%), Chytridiomycota (0.25%, 0.04%) and Rozellomycota (0.12%, 0.006%). The compositions of fungal communities across these three sea regions were found to be vary, which may be attributed to sediment source, geographical distance, latitude and some environmental factors such as the temperature and salinity of bottom water, water depth, total nitrogen, and the ratio of total organic carbon to nitrogen. Among these environmental factors, the temperature of bottom water is the most important driver that governs the distribution patterns of fungal communities across the sampled seas. Our data also suggest that the cold-water mass of the Yellow Sea likely balances competitive relationships between fungal taxa rather than increasing species richness levels.

[Osteoclasts and early bone remodeling after orthodontic micro-implant placement].

PURPOSE: To observe the incidence of osteoclasts during early bone remodeling after orthodontic micro-implant placement. METHODS: Twenty New Zealand rabbits were randomly allotted into 4 groups. One micro-implant was implanted proximal to the epiphyseal plate of the tibia. Animals were sacrificed on day 3, 7, 14 and 28 (n=5). The sequence of histological changes around the micro-implants were evaluated by hematoxylin and eosin (HE) staining. Osteoclasts were identified by TRAP staining. The differences of the number of the osteoclasts among each time point were analyzed by one way ANOVA with SPSS 19.0 software package. RESULTS: After 3 days of implantation, a large number of erythrocytes, inflammatory cells, mesenchymal cells and bone debris were seen at the implant bone interfaces. Few osteoclasts were observed. On day 7, granular woven bone was formed and some osteoclasts were found in the Howship's lacunae. New bone formation and mineralization were apparent on day 14. Meanwhile, large amounts of osteoclasts were found in the latticed woven bone. On day 28, woven trabeculae with lamellate structures connected to lamellar bone and fewer osteoclasts were identified. Semi-quantitative analysis showed that the number of the osteoclasts was at peak on day 14. There were significant differences among each time point (P<0.01). CONCLUSIONS: Osteoclast activity is closely related to bone formation and remodeling after micro-implant insertion.