A regulation mechanism for the promoter region of the pet II operon in Acidithiobacillus ferrooxidans ATCC23270.

Acidithiobacillus ferrooxidans ATCC23270 is a gram-negative and autotrophic bacillus acquiring energy via the oxidation of iron and sulfur. The pet II operon is involved in the sulfur metabolism of A. ferrooxidans. However, the mechanisms that control the expression of the pet II operon are poorly understood. We previously described that the AFE2726 protein is associated with the expression of the pet II operon. Here, we attempted to analyze the involvement of AFE2726 in the regulation of pet II operon expression. First, pEGF recombinant vectors driven by the promotor of the pet II operon, denoted pEGF-pet II, were constructed. Then, DH5α E. coli cultures containing the vector mentioned above were cultivated in Na2S2O3, as this medium substantially enhances the expression of green fluorescent proteins. To examine the regulatory effect of AFE2726 on the pet II operon, the C62/V and C72/V mutants for AFE2726 were constructed in pEGF-pet II vectors using the site-directed deletion method. Compared to pEFG-pet II and pEFG-pet II-Δ-C62/V, pEFG-pet II-Δ-C72/V reduced the expression of green fluorescent proteins dramatically when transformed into DH5α E.coli in Na2S2O3 medium. This suggested that the 72nd cysteine was a crucial residue of the AFE2726 protein, affecting the response of the pet II operon to sodium thiosulfate. Furthermore, the binding site of AFE2726 on the promotor of the pet II operon was identified using the electrophoretic mobility shift assay (EMSA), and it was found to be a 34bp inverted repeat sequence (named IR4), which ranged from -65 to -32. In summary, our results indicated that the AFE2726 protein regulates the pet II operon by binding to the IR4 sequence in its promotor region, whose function is likely affected by Na2S2O3 binding to its Cys72 residue counterpart.

An anti-inflammatory isoflavone from soybean inoculated with a marine fungus Aspergillus terreus C23-3.

A new isoflavone derivative compound 1 (psoralenone) was isolated from soybean inoculated with a marine fungus Aspergillus terreus C23-3, together with seven known compounds including isoflavones 2-6, butyrolactone I (7) and blumenol A (8). Their structures were elucidated by MS, NMR, and ECD. Psoralenone displayed moderate in vitro anti-inflammatory activity in the LPS-induced RAW264.7 cell model. Compound 2 (genistein) showed moderate acetylcholinesterase (AChE) inhibitory activity whereas compounds 2, 5 (biochanin A), 6 (psoralenol), and 7 exhibited potent larvicidal activity against brine shrimp. Compounds 3 (daidzein), 4 (4'-hydroxy-6,7-dimethoxyisoflavone), and 5-7 showed broad-spectrum anti-microbial activity, and compound 7 also showed moderate 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity.

[Sacral nerve magnetic stimulation combined with extracorporeal shockwave for the treatment of type-â ¢B chronic prostatitis].

OBJECTIVE: To investigate the clinical therapeutic effects of sacral nerve magnetic stimulation (SNMS) combined with extracorporeal shockwave (ECSW) in the treatment of type-ⅢB chronic prostatitis. METHODS: This study included 65 cases of type-ⅢB chronic prostatitis treated in Renji Hospital between March 2017 and August 2018. The patients were aged 34.56 + 7.47 years and had an average disease course of 12.95 + 10.73 months. We randomly assigned the patients to an experimental (n = 33) and a control group (n = 32) to be treated by SNMS+ECSW and biofeedback combined with electrical stimulation, respectively, qd alt, 40 minutes once, for a total of 24 times. Before and after 4 and 8 weeks of treatment, we obtained the NIH-CPSI scores, maximum urinary flow rate (Qmax), average urinary flow rate (Qavg), Self-Rating Depression Scale (SDS) scores and Self-Rating Anxiety Scale (SAS) scores of the patients, recorded their adverse reactions and compared the clinical therapeutic effects between the two groups of patients. RESULTS: After treatment, the experimental group showed significant improvement in the pain score, urination score, quality of life (QOL) score and NIH-CPSI total scores in comparison with the baseline (P < 0.05), even more significant after 8 than after 4 weeks of treatment (P < 0.05), and in all the indexes as compared with the control group (P < 0.05). Qmax and Qavg were remarkably improved at 8 weeks (P < 0.05) and so were SDS and SAS scores at 4 and 8 weeks (P < 0.05), even more significantly in the experimental than in the control group (P < 0.05). Among the 33 patients in the experimental group, 25 (75.8%) responded (14 ［42.4%］ cured or with excellent effect), with a significantly higher effectiveness rate than the control group (7［46.9%］， P < 0.01). No obvious adverse events were observed in any of the patients during the treatment. CONCLUSIONS: SNMS+ECSW can effectively improve the clinical symptoms and QOL of the patients with type-ⅢB chronic prostatitis, without causing obvious adverse reactions. Its long-term therapeutic effect, however, remains to be further studied.

Depsidone Derivatives and a Cyclopeptide Produced by Marine Fungus Aspergillus unguis under Chemical Induction and by Its Plasma Induced Mutant.

A new depsidone derivative (1), aspergillusidone G, was isolated from a marine fungus Aspergillus unguis, together with eight known depsidones (2‒9) and a cyclic peptide (10): agonodepside A (2), nornidulin (3), nidulin (4), aspergillusidone F (5), unguinol (6), aspergillusidone C (7), 2-chlorounguinol (8), aspergillusidone A (9), and unguisin A (10). Compounds 1‒4 and 7‒9 were obtained from the plasma induced mutant of this fungus, while 5, 6, and 10 were isolated from the original strain under chemical induction. Their structures were identified using spectroscopic analysis, as well as by comparison with literature data. The HPLC fingerprint analysis indicates that chemical induction and plasma mutagenesis effectively influenced the secondary metabolism, which may be due to their regulation in the key steps in depsidone biosynthesis. In bioassays, compound 9 inhibited acetylcholinesterase (AChE) with IC50 in 56.75 µM. Compounds 1, 5, 7, 8, and 9 showed moderate to strong activity towards different microbes. Compounds 3, 4, and 5 exhibited potent larvicidality against brine shrimp. In docking studies, higher negative CDOCKER interaction energy and richer strong interactions between AChE and 9 explained the greater activity of 9 compared to 1. Chemical induction and plasma mutagenesis can be used as tools to expand the chemodiversity of fungi and obtain useful natural products.

[Overview of traditional Chinese medicine quality evaluation method based on overall research].

The establishment of quality evaluation of traditional Chinese medicine system that not only accords with Chinese medicine function characteristics but also is recognized as international medical circles, is an arduous task in urgent need of solving the current modernization of traditional Chinese medicine in the process of internationalization. It is difficult to evaluate atraditional Chinese medicine by detection of single active components in traditional Chinesemedicinewiththe western medicine quality controlmethod due to the overall effects of traditional Chinese drugs, the components of the overall diversity, targets, and the complexity of the interaction between components of unpredictable make the Long-term since, domestic and foreign scholars continue to explore and put forward a series of quality evaluation of traditional Chinese medicine to promote the development of traditional Chinese medicine. This article summarized the related academic ideas and developments to, providea new thought and perspective for the quality control of traditional Chinese medicine.

[Preparation of novel gypenosides long-circulating liposomes consisted by sphingomyelin and beta-sitosterol modified by PEG].

OBJECTIVE: To explore the feasibility of preparing novel gypenosides long-circulating liposomes with PEG grafted on beta-sitosterol (PEG-Sito). METHOD: Succinicanhydride was adopted to connect beta-sitosterol and PEG 2000. Sphingomyelin and PEG-Sito was used as material to prepare gypenosides long-circulating liposomes by using ethanol injection method. Encapsulation efficiency was determined by using protamine precipitation method. H-NMR was used to verify the synthesis of PEG-Sito, the novel gypenosides long-circulating liposomes were characterized by particle size, zeta potential and atomic force microscope. RESULT: The synthesis of PEG-Sito was verified by 1H-NMR. Encapsulation efficiency of long-circulating liposomes prepared by ethanol injection method was 74.3%, particle size was 288.1 nm, zeta potential was -20.25 mV, the morphology were round observed by AFM. CONCLUSION: The novel gypenosides long-circulating liposomes prepared with PEG-Sito was feasible, it had a high encapsulation efficiency and good morphology.

An efficient flexible graphene-based light-emitting device.

In recent years, flexible light-emitting devices (LEDs) have become the main focus in the field of display technology. Graphene, a two-dimensional layered material, has attracted great interest in LEDs due to its excellent properties. However, there are many problems such as efficiency, lifetime, and flexibility not well solved. Herein, we have successfully prepared a flexible LED using laser-induced reduced graphene oxide (LIRGO). The LIRGO LED achieves a luminescence lifetime of over 60 hours and a wall plug efficiency of up to 1.4% in a vacuum environment of 0.02 Pa. There are many small luminescent spots randomly distributed on 3.5 × 5 mm2 of LIRGO. LIRGO's luminous behavior can be controlled by modifying the supply voltage and laser reduction intensity. We also explore LIRGO's applications by testing it in different packages and customizable bulbs. Furthermore, as an interesting demo, the LIRGO device can be used to mimic constellations with visual shapes. This work demonstrates LIRGO's great potential in many fields, such as flexible and miniature light sources and displays.

Feeble Antipyretic, Analgesic, and Anti-inflammatory Activities were Found with Regular Dose 4'-O-ß-D-Glucosyl-5-O-Methylvisamminol, One of the Conventional Marker Compounds for Quality Evaluation of Radix Saposhnikoviae.

INTRODUCTION: 4'-O-ß-D-glucosyl-5-O-methylvisamminol (GML) is a conventional marker compound for quality control of Radix Saposhnikoviae. Despite that, neither pharmacodynamic or pharmacokinetic information is available with regard to GML. As such, the aim of thisstudy was to assess the conventional evaluation indices for the quality of Radix Saposhnikoviae. MATERIALS AND METHODS: Pyretic animal model, hot plate test, and ear edema model were established to evaluate and compare the antipyretic, analgesic, and anti-inflammatory effect of the chromone derivativescimifugin, prime-O-glucosylcimifugin (PGCN), and GML in Radix Saposhnikoviae. High performance liquid chromatography separation and analysis was used to obtain pharmacokinetic parameters. Simulated gastric fluid and simulated intestinal fluid was used to investigate the metabolite profiles of PGCN and GML in gastrointestinal tract. RESULTS: Cimifugin exerted a marked dose-dependent antipyretic, analgesic, and anti-inflammatory effect, whereas the effects of PGCN were relatively lower. GML had feeble pharmacodynamic effects. Pharmacokinetic study showed that only cimifugin was detected in the plasma sample of cimifugin and PGCN-treated animals, with drug concentration in the former much higher than the latter. No components were traced in the plasma samples from GML-treated rats. Stability study showed that PGCN and GML was predominantly biotransformed into cimifugin and 5-O-methyvisammiol, respectively. The latter was proven to be extremely unstable in liver tissue homogenate and plasma. CONCLUSIONS: A feeble antipyretic, analgesic, and anti-inflammatory activities was observed when GML was orally delivered. Given that Radix Saposhnikoviae extract is generally administered orally, we speculate that this compound might be a nonpharmacolagically active agent in real usage. Thus, it might be unscientific to evaluate the quality of Radix Saposhnikoviae based on the content of GML. SUMMARY: GML-derived cimifugin, which represents the potential pharma codynamic component of Radix Saposhnikoviae chromones, in plasma was almost nil in contrast to cimifugin and PGCN. And thus, feeble antipyretic, analgesic, and anti-inflammatory activities were found with GML. Abbreviations used: AUC:area under concentration-time curve, DNP:2,4-Dinitrophenol, HPLC:high performance liquid chromatography, HPLC-MS:high performance liquid chromatography- mass spectrography, GML:4'-O-ß-D-glucosyl-5-O-methylvisamminol, MVL:5-O-methyvisammiol, PGCN:prime-O-glucosylcimifugin, SGF:alkaline phosphatase. SIF:simulated intestinal fluid.

Polysaccharide enhances Radix Saposhnikoviae efficacy through inhibiting chromones decomposition in intestinal tract.

Vegetative but not reproductive stage of Saposhnikovia divaricate (Turxz.) schischk possesses pharmacological activities. However, our recent study showed that reproductive S. divaricate supplemented with polysaccharide showed evidently elevated pharmacological activities and increased cimifugin content in rat serum. The aims of present study were to assess the influence of polysaccharides on the chromones pharmacological activities in Radix Saposhnikoviae (RS), the dried root of vegetative stage of S. divaricate, and to explore the underlying mechanisms. Only cimifugin was detected in the plasma of chromone treated animals and RS polysaccharide significantly increased the plasma content of cimifugin. It was shown that neither cimifugin absorption nor glycoside components transformation in simulated digestive fluid was affected by RS polysaccharide. However, a significant promotion of transformation of cimifugin to more stable prime-O-glucosylcimifugin (PGCN) by RS polysaccharide, and a protective effect of polysaccharide on chromone components were observed in small intestine solutions. Meanwhile, RS polysaccharide produced a significant elevation of cimifugin and PGCN concentration in vivo. Based on these findings, we concluded that RS polysaccharide could greatly increase the content of cimifugin, which might be related to its degradation-proof effect on cimifugin, via transforming cimifugin to comparatively more stable PGCN and spatial structure protection.

[Clinical study on idiopathic thrombocytopenic purpura treated with Shengxueling Granule].

OBJECTIVE: To observe the clinical effect of Shengxueling Granule (SXLG) in treating idiopathic thrombocytopenic purpura (ITP), and to study its possible mechanism. METHODS: Eighty-six cases of ITP were divided into two groups randomly. Fifty-six cases in the treatment group were treated with SXLG, a traditional Chinese medicine, and 30 cases administered with Western medicine (prednisone) were taken as control. Patients in each group took drugs for three months and were under follow-up observation. RESULTS: In SXLG-treated group, the total effective rate in 3 months was 85.71%, similar to 83.33% in prednisone-treated group (P>0.05), while the total effective rate in 6 months in the SXLG-treated group was 91.07%, higher than 53.33% of the prednisone-treated group (P<0.01), and no obvious side-effects were observed. The patients' bleeding was alleviated or stopped, and their general condition was improved. And the blood platelet count (BPC) was increased, the platelet associated immunoglobulin (PAIg) and interleukin-4 (IL-4) were markedly dropped, the level of natural killer cells activity (NKCA) increased, and the rate of T lymphocyte subsets gradually returned to normal level. Megakaryocytes tended toward maturation on bone marrow smear after SXLG treatment. All differences above were statistically significant. CONCLUSION: SXLG is an effective and safe medicine for ITP. It can regulate the cellular immunity, inhibit the platelet antibody to reduce the destruction of the platelet and to increase the number of platelet, promote the differentiation and maturation of megakaryocyte, facilitate the production and release of platelet, lower the fragility of capillary, and prevent the hemorrhagic tendency.

[Study on the knowledge, behavior and attitude of influenza A (H1N1) among residents in Shuangqiao District, Chengde city].

OBJECTIVE: To understand the the residents' knowledge, behavior and attitude of influenza A (H1N1) in Shuangqiao District of Chengde city, and provide the basis for making proper strategies of health education, prevention and control on influenza A (H1N1). METHODS: 211 residents from 1 community and 1 village of Shuangqiao District were selected to participate the questionnaire interview with multi-stage clustering sampling. RESULTS: 97.6% of the interviewed had received some kind of information on influenza A (H1N1); Total awareness rate of influenza A (H1N1) knowledge was 58.5%, which increased with the level of education and varied among diverse occupations; 48.2% of respondents conceded that their lives was affected by the influenza A ( H1N1) in some degree, and 9% of selected residents believed that there would be a severe pandemicity in this winter, while 7% of respondents didn't consider any form of preventive methods in the future; 78% of respondents expressed their wishes to be vaccinated, but 11.0% of respondents refused to received inoculation explicitly; on the issue of assessment on services provided by governments and health facilities, 93% of respondents expressed their satisfaction hierarchically. Conclusion Residents in Shuangqiao District lacked of comprehensive knowledge of influenza A (H1N1), and some specific health education should be carried out.