Mangiferin prevents hepatocyte epithelial-mesenchymal transition in liver fibrosis via targeting HSP27-mediated JAK2/STAT3 and TGF-ß1/Smad pathway.

Hepatocytes has been confirmed to undergo EMT and can be converted into myofibroblasts during hepatic fibrogenesis. However, the mechanism of hepatocyte EMT regulation in hepatic fibrosis, particularly through HSP27 (human homologue of rodent HSP25), remains unclear. Mangiferin (MAN), a compound extracted from Mangifera indica L, has been reported to attenuate liver injury. This study aimed to investigate the mechanisms underlying HSP27 inhibition and the anti-fibrotic effect of MAN in liver fibrosis. Our results revealed that the expression of HSP27 was remarkably increased in the liver tissues of patients with liver cirrhosis and CCl4 -induced fibrotic rats. However, HSP27 shRNA treatment significantly alleviated fibrosis. Furthermore, MAN was found to inhibit CCl4 - and TGF-ß1-induced liver fibrosis and reduced hepatocyte EMT. More importantly, MAN decreased HSP27 expression to suppress the JAK2/STAT3 pathway, and subsequently blocked TGF-ß1/Smad signaling, which were consistent with its protection against CCl4 -induced EMT and liver fibrosis. Together, these results suggest that HSP27 may play a crucial role in hepatocyte EMT and liver fibrosis by activating JAK2/STAT3 signaling and TGF-ß1/Smad pathway. The suppression of HSP27 expression by MAN may be a novel strategy for attenuating the hepatocyte EMT in liver fibrosis.

Isolation, Phylogenetic and Gephyromycin Metabolites Characterization of New Exopolysaccharides-Bearing Antarctic Actinobacterium from Feces of Emperor Penguin.

A new versatile actinobacterium designated as strain NJES-13 was isolated from the feces of the Antarctic emperor penguin. This new isolate was found to produce two active gephyromycin analogues and bioflocculanting exopolysaccharides (EPS) metabolites. Phylogenetic analysis based on pairwise comparison of 16S rRNA gene sequences showed that strain NJES-13 was closely related to Mobilicoccus pelagius Aji5-31T with a gene similarity of 95.9%, which was lower than the threshold value (98.65%) for novel species delineation. Additional phylogenomic calculations of the average nucleotide identity (ANI, 75.9-79.1%), average amino acid identity (AAI, 52.4-66.9%) and digital DNA-DNA hybridization (dDDH, 18.6-21.9%), along with the constructed phylogenomic tree based on the up-to-date bacterial core gene (UBCG) set from the bacterial genomes, unequivocally separated strain NJES-13 from its close relatives within the family Dermatophilaceae. Hence, it clearly indicated that strain NJES-13 represented a putative new actinobacterial species isolated from the gut microbiota of mammals inhabiting the Antarctic. The obtained complete genome of strain NJES-13 consisted of a circular 3.45 Mb chromosome with a DNA G+C content of 67.0 mol%. Furthering genome mining of strain NJES-13 showed the presence of five biosynthetic gene clusters (BGCs) including one type III PKS responsible for the biosynthesis of the core of gephyromycins, and a series of genes encoding for bacterial EPS biosynthesis. Thus, based on the combined phylogenetic and active metabolites characterization presented in this study, we confidently conclude that strain NJES-13 is a novel, fresh actinobacterial candidate to produce active gephyromycins and microbial bioflocculanting EPS, with potential pharmaceutical, environmental and biotechnological implications.

Sulfitobacter alexandrii sp. nov., a new microalgae growth-promoting bacterium with exopolysaccharides bioflocculanting potential isolated from marine phycosphere.

Marine phycosphere harbors unique cross-kingdom associations with enormous ecological significance in aquatic ecosystems as well as relevance for algal biotechnology industry. During our investigating the microbial composition and bioactivity of marine phycosphere microbiota (PM), a novel lightly yellowish and versatile bacterium designated strain AM1-D1T was isolated from cultivable PM of marine dinoflagellate Alexandrium minutum amtk4 that produces high levels of paralytic shellfish poisoning toxins (PSTs). Strain AM1-D1T demonstrates notable bioflocculanting bioactivity with bacterial exopolysaccharides (EPS), and microalgae growth-promoting (MGP) potential toward its algal host. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain AM1-D1T was affiliated to the members of genus Sulfitobacter within the family Rhodobacteraceae, showing the highest sequence similarity of 97.9% with Sulfitobacter noctilucae NB-68T, and below 97.8% with other type strains. The complete genome of strain AM1-D1T consisted of a circular 3.84-Mb chromosome and five circular plasmids (185, 95, 15, 205 and 348 Kb, respectively) with the G+C content of 64.6%. Low values obtained by phylogenomic calculations on the average nucleotide identity (ANI, 77.2%), average amino acid identity (AAI, 74.7%) and digital DNA-DNA hybridization (dDDH, 18.6%) unequivocally separated strain AM1-D1T from its closest relative. The main polar lipids were identified as phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, diphosphatidylglycerol, one unidentified phospholipid and one unidentified lipid. The predominant fatty acids (> 10%) were C18:1 ω7c, C19:0 cyclo ω8c and C16:0. The respiratory quinone was Q-10. The genome of strain AM1-D1T was predicted to encode series of gene clusters responsible for sulfur oxidation (sox) and utilization of dissolved organic sulfur exometabolites from marine dinoflagellates, taurine (tau) and dimethylsulfoniopropionate (DMSP) (dmd), as well as supplementary vitamin B12 (cob), photosynthesis carotenoids (crt) which are pivotal components during algae-bacteria interactions. Based on the evidences by the polyphasic characterizations, strain AM1-D1T represents a novel species of the genus Sulfitobacter, for which the name Sulfitobacter alexandrii sp. nov. is proposed. The type strain is AM1-D1T (= CCTCC 2017277T = KCTC 62491T).

Anti-inflammatory and neuroprotective effects of natural cordycepin in rotenone-induced PD models through inhibiting Drp1-mediated mitochondrial fission.

Accumulating evidences suggest that inflammation-mediated neurons dysfunction participates in the initial and development of Parkinson's disease (PD), whereas mitochondria have been recently recognized as crucial regulators in NLRP3 inflammasome activation. Cordycepin, a major component of cordyceps militaris, has been shown to possess neuroprotective and anti-inflammatory activity. However, the effects of cordycepin in rotenone-induced PD models and the possible mechanisms are still not fully understood. Here, we observed that motor dysfunction and dopaminergic neurons loss induced by rotenone exposure were ameliorated by cordycepin. Cordycepin also reversed Drp1-mediated aberrant mitochondrial fragmentation through increasing AMPK phosphorylation and maintained normal mitochondrial morphology. Additionally, cordycepin effectively increased adenosine 5'-triphosphate (ATP) content, mitochondrial membrane potential (MMP), and reduced mitochondrial ROS levels, as well as inhibited complex 1 activity. More importantly, cordycepin administration inhibited the expression of NLRP3 inflammasome components and the release of pro-inflammatory cytokine in rotenone-induced rats and cultured neuronal PC12 cells. Moreover, we demonstrated that the activation of NLRP3 inflammasome within neurons could be suppressed by the mitochondrial division inhibitor (Mdivi-1). Collectively, the present study provides evidence that cordycepin exerts neuroprotective effects partially through preventing neural NLRP3 inflammasome activation induced by Drp1-dependent mitochondrial fragmentation in rotenone-injected PD models.

Comparative Proteomic Analysis of Mature Pollen in Triploid and Diploid Populus deltoides.

Ploidy affects plant growth vigor and cell size, but the relative effects of pollen fertility and allergenicity between triploid and diploid have not been systematically examined. Here we performed comparative analyses of fertility, proteome, and abundances of putative allergenic proteins of pollen in triploid poplar 'ZhongHuai1' ('ZH1', triploid) and 'ZhongHuai2' ('ZH2', diploid) generated from the same parents. The mature pollen was sterile in triploid poplar 'ZH1'. By applying two-dimensional gel electrophoresis (2-DE), a total of 72 differentially expressed protein spots (DEPs) were detected in triploid poplar pollen. Among them, 24 upregulated and 43 downregulated proteins were identified in triploid poplar pollen using matrix-assisted laser desorption/ionisation coupled with time of-flight tandem mass spectrometer analysis (MALDI-TOF/TOF MS/MS). The main functions of these DEPs were related with "S-adenosylmethionine metabolism", "actin cytoskeleton organization", or "translational elongation". The infertility of triploid poplar pollen might be related to its abnormal cytoskeletal system. In addition, the abundances of previously identified 28 putative allergenic proteins were compared among three poplar varieties ('ZH1', 'ZH2', and '2KEN8'). Most putative allergenic proteins were downregulated in triploid poplar pollen. This work provides an insight into understanding the protein regulation mechanism of pollen infertility and low allergenicity in triploid poplar, and gives a clue to improving poplar polyploidy breeding and decreasing the pollen allergenicity.

Therapeutic effect of acupuncture combining standard swallowing training for post-stroke dysphagia: A prospective cohort study.

OBJECTIVE: To assess the therapeutic effect of acupuncture combining standard swallowing training for patients with dysphagia after stroke. METHODS: A total of 105 consecutively admitted patients with post-stroke dysphagia in the Affiliated Hospital of Gansu University of Chinese Medicine were included: 50 patients from the Department of Neurology and Rehabilitation received standard swallowing training and acupuncture treatment (acupuncture group); 55 patients from the Department of Neurology received standard swallowing training only (control group). Participants in both groups received 5-day therapy per week for a 4-week period. The primary outcome measures included the scores of Videofluoroscopic Swallow Study (VFSS) and the Standardized Swallowing Assessment (SSA); the secondary outcome measure was the Royal Brisbane Hospital Outcome Measure for Swallowing (RBHOMS), all of which were assessed before and after the 4-week treatment. RESULTS: A total of 98 subjects completed the study (45 in the acupuncture group and 53 in the control group). Significant differences were seen in VFSS, SSA and RBHOMS scores in each group after 4-week treatment as compared with before treatment (P<0.01). Comparison between the groups after 4-week treatment showed that the VFSS P=0.007) and SSA scores (P=0.000) were more significantly improved in the acupuncture group than the control group. However, there was no statistical difference (P=0.710) between the acupuncture and the control groups in RBHOMS scores. CONCLUSIONS: Acupuncture combined with the standard swallowing training was an effective therapy for post-stroke dysphagia, and acupuncture therapy is worth further investigation in the treatment of post-stroke dysphagia.

Effect of acupuncture on inflammatory cytokines expression of spastic cerebral palsy rats.

OBJECTIVE: To To investigate the effect of acupuncture on the tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), C-reactive protein (CRP), nitric oxide synthase (NOS) content and muscular tension of spasticity cerebral palsy rat model. METHODS: The rats with spastic cerebral palsy were randomly divided into the control group, model group and acupuncture group. After successful modeling, the muscular tension and the content of TNF-α, IL-6, CRP, NOS were measured. RESULTS: The serum TNF-α, IL-6, CRP, NOS content were significantly decreased in the acupuncture group (P<0.05). The low and high shear viscosity of whole blood of the acupuncture group were significantly lower than the control group and the model group (P<0.05). The erythrocyte electrophoresis indexes in the acupuncture group were significantly lower than that in the model group and the control group (P<0.05). Acupuncture significantly reduced the muscular tension of spastic cerebral palsy rat and increased the active extent in the paralytic extremity (P<0.05), but it could not be restored to normal level. Compared with the control group, the difference had significant (P<0.05). CONCLUSIONS: Acupuncture treatment can inhibit the release of inflammatory cells after brain injury, then reduce immune injury, relieve muscle spasms and reduce muscular tension.

[Special culture mediums of breeding clones of Dendrobium officinale].

The explants were obtained from stem segments of hybrid combination of Dendrobium officinale germplasms (Zhejiang Yandang Mountain x Yunnan Guangnan). The screened culture mediums for buds induction and multiplication, which were 1/2MS + IBA 1.0 mg x L(-1) +6-BA 1.0 mg x L(-1) and 1/2MS + IBA 1.5 mg x L(-1) +6-BA 0.5 mg x L(-1) respectively, were applied to 159 germplasms of D. officinale from Zhejiang, Yunnan, Guangxi, Hunan, etc. The medium for axillary buds induction had universality with 94.3% induction rate. During buds multiplication, there were significant differences in proliferation effect among germplasms, and three proliferation forms i.e. single bud, multiple buds and protocorm were differentiated from different germplasms. The results showed that different germplasms of D. officinale had specific requirements for culture medium. Therefore, developing special culture medium for breeding clones of D. officinale is urgent and has important application values.

[Studies on agronomic traits of seedlings from different F1 generations of Dendrobium officinale].

OBJECTIVE: To select superior mating combinations from different F1 generations of Dendrobium officinale. METHOD: Twenty-six superior parents from four provenances were selected to make up 26 mating combinations. The agronomic traits of different F1 generation were measured, counted, and analyzed by DPS software. RESULT: There were significant differences between different mating combinations among F1 generations. The variation among full-sib families was significantly higher than that among half-sib families, while the variation inside full-sib families was significantly lower than that inside half-sib families. Twelve agronomic traits were simplified into four principal components, of which the accumulative contribution rate was 84.127 3%. Twenty-six mating combinations have been divided into six groups, plants from the fifth and sixth group grew much better than the others. CONCLUSION: The selection of parents should be emphasized by using hybrid vigor. There were higher genetic gains and superior uniformity inside families when selection and breeding occurs in full-sib families than in half-sib families. Eight superior full-sib families (83 x 34, 66 x 9, 68 x 2, 91 x 69, 66 x 65, 69 x 91, 17 x 66, 66 x 17) have been selected which showed a good agronomic traits of seedlings.

[Field experiment of F1 generation and superior families selection of Dendrobium officinale].

Based on randomized block design of experiment, agronomic traits and yields of 14 F1 generations of Dendrobium officinale were determined. The results showed that the differences in agronomic traits and yields among families were significant, and the hybrid vigor was obvious. Families of 6b x 2a, 9 x 66 and 78 x 69 were selected with the remarkable superiority of yields, agronomic traits and product customization. Correlation analysis between agronomic traits and yields showed that plant height, stem diameter, leaf number, blade length and blade width were all significantly correlated with biological yields and economic yields. Among which, stem diameter, leaf number and blade length were the most significant, and an optimal linear regression model could be established. When the number of shoots was fewer than 4.5, both biological yields and economic yields increased with the increasing number of shoots, but it could not much affect yields when the number of shoots was larger than 4.5. Shoots number, stem diameter and leaf index were basic stability when compared biennial traits to annual, which could be used for early selection.

[Quantitive variation of polysaccharides and alcohol-soluble extracts in F1 generation of Dendrobium officinale].

Using phenol-sulfuric acid method and hot-dip method of alcohol-soluble extracts, the contents of polysaccharides and alcohol-soluble extracts in 11 F1 generations of Dendrobium officinale were determined. The results showed that the polysaccharides contents in samples collected in May and February were 32.89%-43.07% and 25.77%-35.25%, respectively, while the extracts contents were 2.81%-4.85% and 7.90%-17.40%, respectively. They were significantly different among families. The content of polysaccharides in offspring could be significantly improved by hybridization between parents with low and high polysaccharides contents, and the hybrid vigor was obvious. Cross breeding was an effective way for breeding new varieties with higher polysaccharides contents. Harvest time would significantly affect the contents of polysaccharides and alcohol-soluble extracts. The contents of polysaccharides in families collected in May were higher than those of polysaccharides in families collected in February, but the extracts content had the opposite variation. The extents of quantitative variation of polysaccharides and alcohol-soluble extracts were different among families, and each family had its own rules. It would be significant in giving full play to their role as the excellent varieties and increasing effectiveness by studying on the quantitative accumulation regularity of polysaccharides and alcohol-soluble extracts in superior families (varieties) of D. officinale to determine the best harvesting time.

[Clinical research of chloasma treated with the meridian cosmetology and the impacts on estrogen and progestin].

OBJECTIVE: To observe the efficacy difference between meridian cosmetology and western medicine in the treatment of chloasma and explore the impacts of meridian cosmetology on E2 and PRGE. METHODS: Three hundreds cases of chloasma were randomized into a meridian cosmetology group and a western medication group according to the visit sequence, 150 cases in each one. In the meridian cosmetology group, the meridian regulation, acupuncture based on pattern/syndrome differentiation [at the body acupoints such as Qihai (CV 6), Xuehai (SP 10), Zusanli (ST 36), Sanyinjiao (SP 6), Ganshu (BL 18), Pishu (BL 20) and Shenshu (BL 23), etc.] and the local surrounding needling therapy on the chloasma skin lesion were adopted. In the western medication group, the oral administration of Vitamin C and E was prescribed for 3 months. The clinical efficacy was observed for the patients in the two groups. Additionally, 30 cases were collected from the meridian cosmetology group randomly for the detection of estrogen (E2) and progestin (PRGE) before and after treatment. RESULTS: The effective rate in the meridian cosmetology group was 92.6% (126/136), which was superior to 67.0% (75/112) in the western medication group (P < 0.05). For the patients collected from the meridian cosmetology group, as compared with that before treatment, E2 level was decreased (P < 0.01) and PRGE level was increased (P < 0.05). CONCLUSION: The meridian cosmetology achieves the superior efficacy as compared with the western medication group for the chloasma and it can regulate the levels of E2 and PRGE.

[Effects of organic loading rate on the cultivation and characteristic of granular sludge with phosphorus removal].

In an anaerobic/aerobic sequencing batch reactor (SBR), the effect of organic loading rate (OLR) on the formation and phosphorus removal efficiency of granular sludge was investigated under the condition of 1.0-2.0 mg/L of dissolved oxygen (DO) concentration in the period of aerobic feast time in an SBR-cycle. Experiment results showed that the sludge in the SBR was the mixture of a majority of activated sludge floc and a small quantity of round-shaped granular at the OLR of 0.482-0.986 kg/(m3 x d). Granulation was accelerated at the early time of the OLR of 2.218-2.263 kg/(m3 x d), while the morphology of granules changed to an irregular shape on the end of the OLR of 2.218-2.263 kg/(m x d). Then the OLR decreased to 1.113-1.182 kg/(m3 x d), the mature granules were cultivated successfully. The morphology of the mature granules was nearly spherical with a very clear outline. The dimension of granules was 1-2 mm, and the granular sludge had a good settle ability [sludge volume index (SVI) between 20-22 mL/g]. The OLR also had influence on efficiency of phosphorus removal, when the OLR was elevated to 2.218-2.263 kg/(m3 x d), the efficiency of phosphorus removal declined from 99% to 78%, and could not be recovered even if the OLR was decreased to 1.113-1.182 kg/(m3 x d).

[Experiment study on anoxic phosphate accumulation with nitrite].

Biological phosphorus removal with nitrite was studied in an Sequencing Batch Reactor. The results showed that nitrite can be used as electron acceptor in denitrifying phosphorus removal. Feed mode of nitrite had significant influence on denitrifying phosphorus removal. Anoxic phosphorus assimilation rate could be reached 10.44 mg/(g x h) and percentage of anoxic phosphorus assimilation amount was more than 97%. Granular sludge with denitrifying phosphorus removal activity was cultivated in an SBR. The effects of different operational conditions on formation of granular were studied such as COD loading, settling time, HRT and so on. The average dimension of granular was 315 microm. Its water content was at 96.78%-98.14%, specific gravity 1.0064-1.016 , SVI of sludge 25 40 mL/g, COD removal loading could be reached to 1 5 kg/(m3 x d), nitrogen and phosphorus removal efficiency was more than 90%.

Four new cycloartane glycosides from Thalictrum fortunei.

Four new cycloartane glycosides were isolated from the aerial parts of Thalictrum fortunei (Ranunculaceae). The chemical structures of these new glycosides were elucidated as 3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-fucopyranosyl (22S,24Z)-cycloart-24-en-3beta,22,26-triol 26-O-beta-D-glucopyranoside, 3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-fucopyranosyl (22S,24Z)-cycloart-24-en-3beta,22,26-triol 26-O-beta-D-quinovopyranosyl-(1-->6)-beta-D-glucopyranoside, 3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-fucopyranosyl (22S,24Z)-cycloart-24-en-3beta,22,26-triol 26-O-beta-D-xylopyranosyl-(1-->6)-beta-D-glucopyranoside, and 3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-fucopyranosyl (22S,24Z)-cycloart-24-en-3beta,22,26-triol 26-O-alpha-L-arabinopyranosyl-(1-->6)-beta-D-glucopyranoside by extensive NMR methods, HR-ESI-MS, and hydrolysis. This is the first report of (22S,24Z)-3beta,22,26-trihydroxycycloartan-24-ene (thelictogenin A, 5) being glycosylated at C-26.

[Investigation on inhibitory and apoptosis-inducing effects of saponins from Tribulus terrestris on hepatoma cell line BEL-7402].

OBJECTIVE: To investigate the inhibitory and apoptosis-inducing effects of saponins from Tribulus terrestris (STT) on liver cancer cell line BEL-7402. METHOD: MTT, SRB, Wright staining, acridine orange staining, flow cytometry, and Immunofluorescence microscopy were used to evaluate the effects of STT on BEL-7402 cell line. RESULT: SMT had potent inhibitory effect on BEL-7402 cell line in a concentration-dependent manner. BEL-7402 cells exibited typical morphological alteration of apoptosis when sub-G1 peak could be seen. The expression of Bcl-2 was decreased in STT treated cells as compared with untreated control cells. CONCLUSION: STT exerts its cytotoxic effect on BEL-7402 cells by inducing apoptosis.

[Experiment of simultaneous denitrifying phosphorus accumulation].

Denitrifying phosphorus accumulation (DNPA) and the factors affecting it were studied in a SBR with synthetic wastewater. The results showed that the sludge acclimatized under anaerobic/aerobic operation with good phosphorus removal ability, showed DNPA soon when fed nitrate instead of aeration following the anaerobic stage. Anaerobic stage was a vital premise to DNPA. If DNPA sludge was fed with nitrate prior to anaerobic stage, the DNPA would weaken even disappear. When acetate was used as sole carbon resource in the influent and nitrate did not exist in anaerobic, 1 hour of anaerobic time was optimal. NO3- -concentration in the anoxic was one of the factors affecting DNPA. When nitrate concentration was advanced from 5mg/L to 20mg/L, the percentage of DNPA increased from 11.9% to 48.7% under the condition of anaerobic(2h)-anoxic(1h)-aerobic(2h). But when the NO3- -concentration was enhanced upwards of 20mg/L, the efficiency cannot be improved. Induced DNPA did not disappear even though there was aerobic stage following anoxic stage, but the shorter the aerobic stage lasted the higher proportions of phosphorus removal via DNPA to total removal.

[Long term observation in effects of potassium and calcium supplementation on arterial blood pressure and sodium metabolism in adolescents with higher blood pressure].

OBJECTIVE: To investigate the effects of potassium and calcium supplementation in table salt on reduction of arterial blood pressure and sodium metabolism in adolescents with higher blood pressure. METHODS: A single blind placebo-controlled trial was carried out for two years in 220 adolescents with higher blood pressure, aged 18 - 22 years, who were randomly divided into supplementary group (n = 110) and control group (n = 110). Each of the subjects in the supplementary group and their family members was given 10 mmol of potassium and 10 mmol of calcium mixed in their table salt daily for 24 months. RESULTS: Night urinary sodium and potassium excretion increased (urinary Na(+), P < 0.05; urinary K(+), P < 0.01) and blood pressure lowered by 5.3 mm Hg/1.8 mm Hg in average from the baseline in the supplementary group two years after potassium and calcium supplementation, as compared with that in the control group increased by (1.3/1.7) mm Hg. CONCLUSIONS: Adequate supplement of potassium and calcium in daily table salt intake was an effective way to prevent form hypertension and could promote their urinary sodium excretion and reduction of arterial blood pressure in adolescents with higher blood pressure.