Alginate oligosaccharide assimilation by gut microorganisms and the potential role in gut inflammation alleviation.

Dietary fiber metabolism by gut microorganisms plays important roles in host physiology and health. Alginate, the major dietary fiber of daily diet seaweeds, is drawing more attention because of multiple biological activities. To advance the understanding of alginate assimilation mechanism in the gut, we show the presence of unsaturated alginate oligosaccharides (uAOS)-specific alginate utilization loci (AUL) in human gut microbiome. As a representative example, a working model of the AUL from the gut microorganism Bacteroides clarus was reconstructed from biochemistry and transcriptome data. The fermentation of resulting monosaccharides through Entner-Doudoroff pathway tunes the metabolism of short-chain fatty acids and amino acids. Furthermore, we show that uAOS feeding protects the mice against dextran sulfate sodium-induced acute colitis probably by remodeling gut microbiota and metabolome. IMPORTANCE: Alginate has been included in traditional Chinese medicine and daily diet for centuries. Recently discovered biological activities suggested that alginate-derived alginate oligosaccharides (AOS) might be an active ingredient in traditional Chinese medicine, but how these AOS are metabolized in the gut and how it affects health need more information. The study on the working mechanism of alginate utilization loci (AUL) by the gut microorganism uncovers the role of unsaturated alginate oligosaccharides (uAOS) assimilation in tuning short-chain fatty acids and amino acids metabolism and demonstrates that uAOS metabolism by gut microorganisms results in a variation of cell metabolites, which potentially contributes to the physiology and health of gut.

Antiperspirant effects and mechanism investigation of Mulisan decoction in rats based on plasma metabolomics.

CONTEXT: Mulisan decoction (MLS) is a classic formula of traditional Chinese medicine for treating hyperhidrosis. The mechanism remains unclear. OBJECTIVE: To investigate the antiperspirant effect and underlying mechanisms of MLS. MATERIALS AND METHODS: Fifty rats were divided into control, model, and three doses of MLS intervention groups (n = 10). Rats except for control group were induced diseases features of the applicable scope of MLS via i.p. reserpine (0.5 mg/kg/d) for 10 days. From day 11, MLS groups were administrated orally MLS at 0.6, 3, and 15 g/kg once a day for 14 days, respectively. After the last administration, sweating was induced in all rats via s.c. pilocarpine (25 mg/kg), the right hind foot of rats was stained, and sweat point numbers were observed. Rat serum was collected to detect IL-2, IL-6, IFN-γ, and TNF-α. Rat plasma was collected for endogenous metabolite analysis via UPLC-QE-Focus-MS. RESULTS: Rats treated with MLS presented a significant decrease in sweat point numbers (13.5%), increase in body weight (13.2%), and promotion in the balance of Th1/Th2 cytokine ratio via increasing IL-2 (38.3%), IFN-γ (20.1%), and TNF-α (22.0%) and decreasing IL-6 (24.7%) compared with the model group (p < 0.05). Plasma metabolomics disclosed 15 potential biomarkers related to model rats, of which two could be significantly reversed by MLS (p < 0.05). The involved pathways were pantothenate and CoA biosynthesis, and porphyrin metabolism. CONCLUSIONS: MLS demonstrated a good antiperspirant effect and metabolism improvement. These findings inspire more clinical study validation on immune improvement and antiperspirant effect.

Structural basis for the exolytic activity of polysaccharide lyase family 6 alginate lyase BcAlyPL6 from human gut microbe Bacteroides clarus.

Alginate is the structural polysaccharide of the cell wall of brown algae, which is an important carbon source for marine life. The depolymerization of alginate is dependent on alginate lyases. Recent studies showed that the alginate utilization ability had been obtained by human gut microbes. In contrast to the great number of studies on alginate lyases from marine/soil organisms, studies on alginate lyases from gut microbes are still limited. Here, the structure of a polysaccharide lyase family 6 (PL6) alginate lyase from human gut microbe Bacteroides clarus was solved by X-ray crystallography, which represents the cluster of two-domain PL6 alginate lyases from Bacteroidetes. Similar with the two-domain alginate lyase AlyGC originated from marine bacterium, both the N terminal domain (NTD) and C terminal domain (CTD) of BcAlyPL6 show right-handed parallel ß-helix fold. However, unlike AlyGC, which forms a homodimer, BcAlyPL6 functions as a monomer. Biochemical analysis indicates that the substrate binding affinity is mainly contributed by the NTD while the CTD of BcAlyPL6 is involved in the formation of -1 subsite, which is essential for substrate turnover rate. Furthermore, CTD is involved in shaping a closed catalytic pocket, and deletion of it leads to increased activity towards highly polymerized substrate. Structure comparison of PL6 family alginate lyases implies that the linkers of two-domain alginate lyases might have evolutionary relationship with the N/C terminal extension of single-domain lyases.

Biofuel and chemical production from carbon one industry flux gas by acetogenic bacteria.

Carbon one industry flux gas generated from fossil fuels, various industrial and domestic waste, as well as lignocellulosic biomass provides an innovative raw material to lead the sustainable development. Through the chemical and biological processing, the gas mixture composed of CO, CO2, and H2, also termed as syngas, is converted to biofuels and high-value chemicals. Here, the syngas fermentation process is elaborated to provide an overview. Sources of syngas are summarized and the influences of impurities on biological fermentation are exhibited. Acetogens and carboxydotrophs are the two main clusters of syngas utilizing microorganisms, their essential characters are presented, especially the energy metabolic scheme with CO, CO2, and H2. Synthetic biology techniques and microcompartment regulation are further discussed and proposed to create a high-efficiency cell factory. Moreover, the influencing factors in fermentation and products in carboxylic acids, alcohols, and others such like polyhydroxyalkanoate and poly-3-hydroxybutyrate are addressed. Biological fermentation from carbon one industry flux gas is a promising alternative, the latest scientific advances are expatiated hoping to inspire more creative transformation.

Sleep quality and cognitive impairment in older Chinese: Guangzhou Biobank Cohort Study.

BACKGROUND: evidence concerning the relationship between sleep quality and cognitive impairment is limited and inconsistent. OBJECTIVE: to examine the association of sleep quality with memory impairment and poor cognitive function in a large sample of older Chinese. METHODS: 15,246 participants aged 50+ years of the Guangzhou Biobank Cohort Study who attended the second physical examination from 2008 to 2012 were included. Sleep quality was assessed using Pittsburgh Sleep Quality Index (PSQI), and cognitive performance was assessed using both Delayed Word Recall Test (DWRT) and Mini-Mental State Examination (MMSE). Memory impairment was defined by DWRT score < 4 and poor cognitive function by MMSE score < 25. RESULTS: after adjusting for potential confounders, lower habitual sleep efficiency was associated with a higher risk of memory impairment and poor cognitive function with a dose-response pattern (both P for trend <0.001). The adjusted odds ratio (OR, 95% confidence interval (CI)) for poor cognitive function in those with the sleep efficiency of 75-85%, 65-75% and <65%, versus ≥85%, was 1.31 (1.12-1.53), 1.41 (1.16-1.73) and 1.33 (1.09-1.63), respectively. No association of the global PSQI score with memory impairment or poor cognitive function was found. CONCLUSIONS: in older Chinese people, lower habitual sleep efficiency was associated with a higher risk of memory impairment and poorer cognitive function.

Processive Degradation of Crystalline Cellulose by a Multimodular Endoglucanase via a Wirewalking Mode.

Processive hydrolysis of crystalline cellulose by cellulases is a critical step for lignocellulose deconstruction. The classic Trichoderma reesei exoglucanase TrCel7A, which has a closed active-site tunnel, starts each processive run by threading the tunnel with a cellulose chain. Loop regions are necessary for tunnel conformation, resulting in weak thermostability of fungal exoglucanases. However, endoglucanase CcCel9A, from the thermophilic bacterium Clostridium cellulosi, comprises a glycoside hydrolase (GH) family 9 module with an open cleft and five carbohydrate-binding modules (CBMs) and hydrolyzes crystalline cellulose processively. How CcCel9A and other similar GH9 enzymes bind to the smooth surface of crystalline cellulose to achieve processivity is still unknown. Our results demonstrate that the C-terminal CBM3b and three CBMX2s enhance productive adsorption to cellulose, while the CBM3c adjacent to the GH9 is tightly bound to 11 glucosyl units, thereby extending the catalytic cleft to 17 subsites, which facilitates decrystallization by forming a supramodular binding surface. In the open cleft, the strong interaction forces between substrate-binding subsites and glucosyl rings enable cleavage of the hydrogen bonds and extraction of a single cellulose chain. In addition, subsite -4 is capable of drawing the chain to its favored location. Cellotetraose is released from the open cleft as the initial product to achieve high processivity, which is further hydrolyzed to cellotriose, cellobiose and glucose by the catalytic cleft of the endoglucanase. On this basis, we propose a wirewalking mode for processive degradation of crystalline cellulose by an endoglucanase, which provides insights for rational design of industrial cellulases.

Structural insights into the substrate specificity of a glycoside hydrolase family 5 lichenase from Caldicellulosiruptor sp. F32.

Glycoside hydrolase (GH) family 5 is one of the largest GH families with various GH activities including lichenase, but the structural basis of the GH5 lichenase activity is still unknown. A novel thermostable lichenase F32EG5 belonging to GH5 was identified from an extremely thermophilic bacterium Caldicellulosiruptor sp. F32. F32EG5 is a bi-functional cellulose and a lichenan-degrading enzyme, and exhibited a high activity on ß-1,3-1,4-glucan but side activity on cellulose. Thin-layer chromatography and NMR analyses indicated that F32EG5 cleaved the ß-1,4 linkage or the ß-1,3 linkage while a 4-O-substitued glucose residue linked to a glucose residue through a ß-1,3 linkage, which is completely different from extensively studied GH16 lichenase that catalyses strict endo-hydrolysis of the ß-1,4-glycosidic linkage adjacent to a 3-O-substitued glucose residue in the mixed-linked ß-glucans. The crystal structure of F32EG5 was determined to 2.8 Šresolution, and the crystal structure of the complex of F32EG5 E193Q mutant and cellotetraose was determined to 1.7 Šresolution, which revealed that the exit subsites of substrate-binding sites contribute to both thermostability and substrate specificity of F32EG5. The sugar chain showed a sharp bend in the complex structure, suggesting that a substrate cleft fitting to the bent sugar chains in lichenan is a common feature of GH5 lichenases. The mechanism of thermostability and substrate selectivity of F32EG5 was further demonstrated by molecular dynamics simulation and site-directed mutagenesis. These results provide biochemical and structural insights into thermostability and substrate selectivity of GH5 lichenases, which have potential in industrial processes.

Characterization of a thermostable endo-1,3(4)-ß-glucanase from Caldicellulosiruptor sp. strain F32 and its application for yeast lysis.

ß-1,3-Glucans, important structural components of cell wall or nutritional components of the endosperm, are extensively found in bacteria, fungi, yeast, algae, and plants. The structural complexity of ß-1,3-glucans implies that the enzymatic depolymerization of polysaccharides needs combined activities of distinct enzymes. In this study, Lam16A-GH, the catalytic module of a putative glycoside hydrolase (GH) family 16 laminarinase/lichenase from thermophilic bacterium Caldicellulosiruptor sp. F32, was purified and characterized through heterologous expression in Escherichia coli. Lam16A-GH can hydrolyze both ß-1,3-glucan (laminarin) and ß-1,3-1,4-glucan (barley ß-glucan) revealed by analysis of the products of polysaccharide degradation using thin-layer chromatography (TLC). The time required for the loss of 50 % of its activity is 45 h under the optimal condition of 75 °C and pH 6.5. Oligosaccharides degradation assay indicated that Lam16A-GH can catalyze endo-hydrolysis of the ß-1,4 glycosidic linkage adjacent to a 3-O-substituted glucosyl residue in the mixed linked ß-glucans, as well as the ß-1,3 linkage. The survival rate of Saccharomyces cerevisiae cells depends on the addition of Lam16A-GH, and the cytoplasm protein was released from the apparently deconstructed yeast cells. These results indicate that the bi-functional thermostable Lam16A-GH exhibits unique enzymatic properties and potential for yeast lysis.

UHPLC UHD Q-TOF MS/MS analysis of the impact of sulfur fumigation on the chemical profile of Codonopsis Radix (Dangshen).

Over recent decades sulfur fumigation has been becoming abused in processing some freshly harvested Chinese medicinal herbs, although it is questioned whether sulfur fumigation can result in changes in efficacy and safety of the herbs. One of the herbs commonly processed by sulfur fumigation is Codonopsis Radix (Dangshen). A report showed that lobetyolin content in sulfur-fumigated Dangshen was lower than in air-dried Dangshen. Whereas there is no investigation designed to compare the chemical profiles of the sulfur-fumigated Dangshen and the air-dried Dangshen. In the present study, a rapid and versatile ultra-high-performance liquid chromatography coupled with ultra-high resolution quadrupole time-of-flight mass spectrometry (UHPLC UHD Q-TOF MS/MS) method was developed for comprehensive analysis of the chemical profiles of sulfur-fumigated and air-dried Dangshen samples. Principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) demonstrated that there were significant chemical differences between sulfur-fumigated and air-dried Dangshen samples. Among the changed components, 57 compounds were identified, in which 15 sulfur-containing compounds were detected only in sulfur-fumigated samples. The established methods were successfully applied to discriminate sulfur-fumigated Dangshen among commercial samples. Whether the chemical changes caused by sulfur fumigation affect the clinical efficacy and safety of Dangshen needs to be further investigated.

Engineered acetogenic bacteria as microbial cell factory for diversified biochemicals.

Acetogenic bacteria (acetogens) are a class of microorganisms with conserved Wood-Ljungdahl pathway that can utilize CO and CO2/H2 as carbon source for autotrophic growth and convert these substrates to acetate and ethanol. Acetogens have great potential for the sustainable production of biofuels and bulk biochemicals using C1 gases (CO and CO2) from industrial syngas and waste gases, which play an important role in achieving carbon neutrality. In recent years, with the development and improvement of gene editing methods, the metabolic engineering of acetogens is making rapid progress. With introduction of heterogeneous metabolic pathways, acetogens can improve the production capacity of native products or obtain the ability to synthesize non-native products. This paper reviews the recent application of metabolic engineering in acetogens. In addition, the challenges of metabolic engineering in acetogens are indicated, and strategies to address these challenges are also discussed.

Profiling and characterization of volatile components from non-fumigated and sulfur-fumigated Flos Lonicerae Japonicae using comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry coupled with chemical group separation.

Flos Lonicerae Japonicae (FLJ) is a popular herb used for many centuries in Traditional Chinese Medicine as a treatment of fever and inflammation. Non-fumigated processing of FLJ has been the traditional approach used in post-harvest preparation of the commodity for commercial use. However, in recent years, natural drying processing of FLJ has been replaced by sulfur-fumigation for efficiency and pest control. Sulfur-fumigation can induce changes in the volatile compounds of the herb, altering its medicinal properties. A comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC×GC-TOF/MS) method was established for the resolution and determination of volatile components in non-fumigated and sulfur-fumigated FLJ. In this paper, analysis of the volatile oils in non-fumigated and sulfur-fumigated (including lab-prepared sulfur-fumigated and industrial sulfur-fumigated) FLJ was performed using GC×GC-TOF/MS. Seventy-three representative volatile components were identified, including furans, alkalies, acids, aldehydes, ketones, alcohols, terpenes, esters, and others, as the main components of FLJ volatile oils. The proposed method was successfully applied for rapid and accurate quality evaluation of FLJ and its related medicinal materials and preparations.

[Effect of Twin-block in adolescents with mandibular retrusion and normal anterior overjet].

PURPOSE: To investigate the therapeutic effect of modified fixed Twin-block on patients whose mandible was retruded and anterior overjet was normal. METHODS: Thirty-six adolescents with mandibular retrusion and normal anterior overjet were selected. The goal of occlusal reconstruction was to move the soft tissue pogonion forward to the zero meridian(0°). Fixed Twin-block combined with the mini-implants on the mandibular buccal shelf was used to promote mandibular growth. After 12 months, the appliances were removed step by step to observe the stability of the jaw relationship. Fixed orthodontic treatment was performed to achieve ideal occlusion and coordinated profile. SPSS 13.0 software package was used for Student's t test. RESULTS: The length of mandibular body increased significantly which facilitated the forward movement of the soft tissue pogonion, and the profile changed from convex to straight. Comparison of the results before and after Twin-block treatment showed that Co-Gn, SNB, U1-SN, U6-0°, U1-0°, U1-L1, anterior overbite, anterior overjet, Pog's-0°, L1-0° and lower anterior facial height changed significantly (P＜0.05). Comparison of the results before and after fixed orthodontic treatment showed that PP-MP, SN-MP, U6-0°, IMPA, L1-0°,U1-SN, U1-L1, anterior overbite and overjet changed significantly(P＜0.05). CONCLUSIONS: Modified fixed Twin-block combined with mini-implants on the mandibular buccal shelf can effectively stimulate the mandibular growth and improve the lateral profile of patients with mandibular retrusion and normal overjet.

[Study on HPLC fingerprints of Chrysanthemi Flos before and after sulfur-fumigation].

OBJECTIVE: To evaluate the quality of Chrysanthemi Flos before and after sulfur-fumigation by analysis of HPLC fingerprints. METHODS: RP-HPLC method was applied to establish the chromatographic fingerprint. The separation was performed on a Waters C, (250 mm x 4.6 mm, 5 microm) column with a gradient elution composed of acetonitrile-aqueous solution containing 0.5% phosphoric acid. The column temperature was 35 degrees C, the detective wavelength was set at 268 nm. RESULTS: Eleven batches of Chrysanthemi Flos samples composed of both sun-dried and sulfur-fumigated were analyzed and evaluated respectively. The method of clustering analysis was used, and the chromatographic fingerprint common pattern was finally established. CONCLUSION: The HPLC fingerprint method is reliable and suitable for quality evaluation of Chrysanthemi Flos before and after sulfur-fumigation.

[Clinical efficacy of the combination of miniscrew with clear aligner in controlling the roller coaster effect].

PURPOSE: To investigate the clinical efficacy of miniscrew in controlling the roller coaster effect in long-distance space closure with clear aligner. METHODS: Sixteen adult patients with bimaxillary protrusion were selected. All cases were treated with clear aligner and their four first premolars were extracted. G6 was designed in all cases. In the experimental group, four miniscrews were inserted at the beginning of treatment to control the teeth in three dimensions; while in the control group, the miniscrews were not applied until obvious roller coaster effect appeared. Paired t test was used to analyze the data with Graphpad Prism 6.0 software package. RESULTS: All the extraction space was closed successfully and the lateral profile was improved significantly. The torque of the upper incisor was under well control and no roller coaster effect happened in the experimental group; while in the control group, the lingual inclination of the upper incisors and open bite of the posterior teeth developed(P＜0.05). After the miniscrews intrusion of the upper anterior teeth, the occlusion relationship improved significantly. CONCLUSIONS: Miniscrews effectively prevents and eliminates the roller coaster effect in the process of long-distance space closure with clear aligner.

[Consistency of the landmark identification on two types of cephalograms from cone-beam CT].

PURPOSE: To study the landmark consistency on two types of cephalograms reconstructed from the data of cone-beam CT(CBCT). METHODS: CBCT data of 56 patients without teeth missing were uploaded into Dolphin software and transferred to the cephalograms. In the control group, the right half projection was taken while in the experimental group, the images achieved after 6 individual projections were managed to achieve the synthesized cephalograms. One experienced orthodontists traced the 23 frequently-used landmarks and the consistency was compared by means of Hotelling T2 test with SPSS 13.0 software package. RESULTS: The tracing of anterior nasal ridge, posterior nasal ridge, upper incisor root tip, lower incisor edge, lower incisor root tip and condyle showed significant difference(P<0.05). CONCLUSIONS: The synthesized cephalograms in the experimental group show the anatomical structure more precisely which is more beneficial to landmark identification.

[Clinical study of the effect of fixed Twin-block on adolescent skeletal Class â ¡ malocclusion].

PURPOSE: To evaluate the retention ability of fixed Twin-block appliance and its clinical effect on adolescent skeletal ClassⅡmalocclusion. METHODS: Twenty-six skeletal ClassⅡdivision 1 adolescents (M:12, F: 14; age:11~13 years, average: 11.8 years) were chosen and fixed Twin-block appliance was used to guide the mandibular protrusion for one year. Before and after treatment, cephalometric films were taken to observe the skeletal, dental and soft tissue changes. Graphpad Prism 6.0 software was used for Student's t test. RESULTS: There was no loosening or destruction of the appliance during the treatment and the profile of all cases was improved significantly. The indexes that showed significant difference(P<0.05) included the mandibular length and position (Co-Gn, SNB, ANB, Pog-VL, Pos-VL) , the inclination and position of the maxillary incisors(U1-VL, U1-SN), the sagittal position of the mandibular dentition(L1-VL, L6-VL). The indexes that showed insignificant difference(P>0.05) included the mandibular plane(MP-SN), the length and position of the maxilla (SNA, A-VL) , the vertical position of the maxillary incisor(U1-HL), the position of the maxillary posterior teeth(U6-VL, U6-HL), the mandibular incisor inclination(IMPA) and the vertical position of the mandibular dentition(L6-MP,L1-MP). CONCLUSIONS: Fixed Twin-block appliance can enhance the mandibular anchorage, effectively promote the mandibular growth and improve the facial profile.

Production of enterodiol from defatted flaxseeds through biotransformation by human intestinal bacteria.

BACKGROUND: The effects of enterolignans, e.g., enterodiol (END) and particularly its oxidation product, enterolactone (ENL), on prevention of hormone-dependent diseases, such as osteoporosis, cardiovascular diseases, hyperlipemia, breast cancer, colon cancer, prostate cancer and menopausal syndrome, have attracted much attention. To date, the main way to obtain END and ENL is chemical synthesis, which is expensive and inevitably leads to environmental pollution. To explore a more economic and eco-friendly production method, we explored biotransformation of enterolignans from precursors contained in defatted flaxseeds by human intestinal bacteria. RESULTS: We cultured fecal specimens from healthy young adults in media containing defatted flaxseeds and detected END from the culture supernatant. Following selection through successive subcultures of the fecal microbiota with defatted flaxseeds as the only carbon source, we obtained a bacterial consortium, designated as END-49, which contained the smallest number of bacterial types still capable of metabolizing defatted flaxseeds to produce END. Based on analysis with pulsed field gel electrophoresis, END-49 was found to consist of five genomically distinct bacterial lineages, designated Group I-V, with Group I strains dominating the culture. None of the individual Group I-V strains produced END, demonstrating that the biotransformation of substrates in defatted flaxseeds into END is a joint work by different members of the END-49 bacterial consortium. Interestingly, Group I strains produced secoisolariciresinol, an important intermediate of END production; 16S rRNA analysis of one Group I strain established its close relatedness with Klebsiella. Genomic analysis is under way to identify all members in END-49 involved in the biotransformation and the actual pathway leading to END-production. CONCLUSION: Biotransformation is a very economic, efficient and environmentally friendly way of mass-producing enterodiol from defatted flaxseeds.

Haimufang decoction, a Chinese medicine formula for lung cancer, arrests cell cycle, stimulates apoptosis in NCI-H1975 cells, and induces M1 polarization in RAW 264.7 macrophage cells.

BACKGROUND: Lung cancer has the highest morbidity and mortality in the world and novel treatment strategies are still needed. Haimufang decoction (HMF) is a patented clinical prescription of traditional Chinese medicine for lung cancer treatment. HMF is composed of four herbs and has been applied clinically in advanced cancer patients. However, its therapeutic mechanisms are still unclear. This study aims to elucidate the possible mechanisms of HMF for the treatment of lung cancer. METHODS: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay was applied for evaluating the proliferative effect of HMF in lung cancer cells and monocyte macrophage RAW264.7 cells. Flow cytometer was used to detect the effects of HMF on cell cycle and apoptosis, and western blotting was employed to explore the potential apoptotic mechanisms of HMF on lung cancer cells. For immunomodulatory effect, co-culture system was used to detect the activation of macrophage RAW264.7 cells when treated with HMF, and neutral red assay was used to measure the effect of HMF on the phagocytosis of the activated macrophages. Enzyme linked immunosorbent assay, flow cytometer, and immunofluorescence staining method were employed for the investigation on the underlying mechanisms of the immunomodulatory effect on RAW264.7 induced by HMF. RESULTS: HMF inhibited the proliferation, induced S phase cell cycle arrest, and stimulated apoptosis in lung cancer NCI-H1975 cells, while had negligible cytotoxicity on macrophage RAW264.7 cells. Moreover, HMF could activate macrophage RAW264.7 cells and promote the inhibition activity of RAW264.7 cells against lung cancer cells. And also, HMF activated macrophages and increased their phagocytic activity in a concentration-dependent manner. HMF increased the expression of macrophage activation marker CD40, the level of nitric oxide, the generation of intracellular reactive oxygen species, as well as M1 macrophages cytokines including tumor necrosis factor-α, interleukin-1ß, interleukin 12 p70, and interleukin 6. Further investigation showed that HMF induced M1 but not M2 phenotype polarization in RAW264.7 cells. CONCLUSIONS: HMF can mainly exert anticancer activity via (1) cytotoxicity to human lung cancer cells by proliferation inhibition, cell cycle arrest, and apoptosis induction; and also via (2) immunomodulation via macrophage cells activation and M1 phenotype polarization induction.

Expressions of cardiac sympathetic norepinephrine transporter and beta1-adrenergic receptor decreased in aged rats.

Evidence suggests that the deterioration of communication between the sympathetic nervous system and cardiovascular system always accompanies the aging of human and animals. Cardiac sympathetic norepinephrine (NE) transporter (NET) on presynaptic membrane is a predominant component to eliminate released NE in the synaptic cleft and maintains the sensitivity of the beta-adrenergic receptor (beta-AR). In the present study, we investigated NET and beta1-AR mRNA levels and sympathetic nerve density in cardiac sympathetic ganglion and left ventricular myocardium in 2- and 16-month-old rats with Northern blot analysis and immunohistochemistry. The expression levels of NET mRNA, NET protein and beta1-AR mRNA in the ganglia or myocardia of 16-month-old rats were markedly reduced by 67%, 26%, and 43%, respectively, in comparison with those in 2-month-old rats. Our results also show that aging induces a strong decrease of the catecholaminergic nerve fiber density.

[Investigate of three-dimensional power Doppler ultrasound used in placental perfusion assessment in pre-eclampsia pregnancies].

OBJECTIVE: To investigate the placenta perfusion in pre-eclampsia by three-dimensional power Doppler ultrasound and analysing the clinical outcome. METHOD: 80 cases of normal and pre-eclampsia pregnant women from July 2007 to May 2008 in Beijing Obstetrics and Gynecology Hospital were studied. The control group of 36 cases, pre-eclampsia were divided into three groups: a total of 44 cases, mild pre-eclampsia group (9 cases), severe pre-eclampsia group (26 cases) and chronic hypertension with pre-eclampsia group (9 cases). The placental vascular index (VI), flow index (FI), blood vessels and blood flow index (VFI) by three-dimensional Doppler histogram and the umbilical blood flow systolic to diastolic (S/D) by color Doppler flow imaging were calculated and the gestational age after delivery, birth weight, placental weight after birth were recorded respectively. RESULTS: (1) VI, FI, VFI and umbilical blood flow S/D value: 6.3+/-2.9, 38.6+/-4.4, 2.7+/-1.3, 2.5+/-0.6 in normal group; 5.7+/-3.8, 36.3+/-7.2, 2.4+/-2.0, 2.4+/-0.3 in mild pre-eclampsia group; 3.0+/-2.4, 31.7+/-5.0, 1.1+/-1.0, 2.9+/-1.3 in severe pre-eclampsia group; 2.2+/-1.6, 26.1+/-4.4, 0.8+/-0.6, 3.1+/-1.6 in chronic hypertension with pre-eclampsia group. Placenta of normal group and mild preeclampsia group VI, FI, VFI were no significant difference (P>0.05), placental VI, FI, VFI of severe preeclampsia group and chronic hypertension with severe pre-eclampsia group were significantly lower than the normal group (P<0.01) and than mild pre-eclampsia group (P<0.05). The umbilical blood flows were not obvious difference among four groups (P>0.05); (2) Gestational age after birth, birth weight, low newborn weight rate and placental weight: (38.7+/-1.5) weeks, (3280+/-520) g, 3%, (568+/-141) g in normal group; (37.9+/-1.0) weeks, (2971+/-265) g, 0, (576+/-98) g in mild pre-eclampsia group; (33.2+/-2.6) weeks, (1820+/-737) g, 58%, (458+/-154) g in severe pre-eclampsia group; (32.6+/-2.6) weeks, (1497+/-533) g, 7/9, (396+/-141) g in chronic hypertension with pre-eclampsia group. The normal group and mild pre-eclampsia group in gestational age after birth, birth weight, low newborn weight rate and placental weight were not significant difference (P>0.05); severe pre-eclampsia and chronic hypertension with severe pre-eclampsia group in them were significantly lower than the normal group (P<0.01) and than mild pre-eclampsia group (P<0.05). CONCLUSIONS: (1) Placental blood flow perfusion of the severe pre-eclampsia and pre-eclampsia with chronic hypertension of pregnancy decreased resulting in clinically lower placental weight, birth weight and gestational age at delivery, but there were no obvious differences in umbilical blood flow S/D values. (2) The investigation was helpful to clinical diagnosis in the placenta perfusion of pre-eclampsia by three-dimensional power Doppler ultrasound.