Performance evaluation of rhamnolipid biosurfactant produced by Pseudomonas aeruginosa and its effect on marine oil-spill remediation.

This study aimed to reveal that the effect of biosurfactant on the dispersion and degradation of crude oil. Whole genome analysis showed that Pseudomonas aeruginosa GB-3 contained abundant genes involved in biosurfactant synthesis and metabolic processes and had the potential to degrade oil. The biosurfactant produced by strain GB-3 was screened by various methods. The results showed that the surface tension reduction activity was 28.6 mN·m-1 and emulsification stability was exhibited at different pH, salinity and temperature. The biosurfactant was identified as rhamnolipid by LC-MS and FTIR. The fermentation conditions of strain GB-3 were optimized by response surface methodology, finally the optimal system (carbon source: glucose, nitrogen source: ammonium sulfate, C/N ratio:16:1, pH: 7, temperature: 30-35 °C) was determined. Compared with the initial fermentation, the yield of biosurfactant increased by 4.4 times after optimization. In addition, rhamnolipid biosurfactant as a dispersant could make the dispersion of crude oil reach 38% within seven days, which enhanced the bioavailability of crude oil. As a biostimulant, it could also improve the activity of indigenous microorganism and increase the degradation rate of crude oil by 10-15%. This study suggested that rhamnolipid biosurfactant had application prospect in bioremediation of marine oil-spill.

Pollution characteristics of groundwater in an agricultural hormone-contaminated site and implementation of Fenton oxidation process.

The groundwater polluted by an agricultural hormone site was taken as the research object, and a total of 7 groundwater samples were collected at different locations in the plant. The main pollutants in the research area were determined to be extractable petroleum hydrocarbons (C10-C40); 1,2-dichloroethane; 1,1,2-trichloroethane; carbon tetrachloride; vinyl chloride, and chloroform; the maximum content of these pollutants can reach 271 mg/L, 1.68 × 107 µg/L, 1.56 × 104 µg/L, 9.53 × 104 µg/L, 6.58 × 104 µg/L, and 4.81 × 104 µg/L, respectively. Aiming at the problems of groundwater pollution in this area, two sets of oxidation experiments have been carried out. The addition of NaHSO3 modified Fenton oxidation system was used in this contaminated water, which enhanced (2.2 ~ 46.7%) chemical oxygen demand (COD) removal rate. The highest removal rate of extractable petroleum hydrocarbons (C10-C40) can reach 99%. And the degradation rate of chlorinated hydrocarbon pollutants can reach 99% to 100%, which almost achieved the purpose of complete removal. In the NaHSO3 modified Fenton oxidation system, the addition of NaHSO3 accelerates the cycle of Fe3+/Fe2+ and ensures the continuous existence of Fe2+ in the reaction system, thereby producing more ·OH and further oxidizing and degrading organic pollutants. Our work has provided important insights for this economically important treatment of this type water body and laid the foundation for the engineering of this method.

Degradation characteristics of crude oil by a consortium of bacteria in the existence of chlorophenol.

In order to enhance the degradation effect of microorganisms on crude oil in the existence of chlorophenol compounds, oil-degrading bacteria C4 (Alcaligenes faecails), C5 (Bacillus sp.) and 2,4-dichlorophenol (2,4-DCP) degrading bacteria L3 (Bacillus marisflavi), L4 (Bacillus aquimaris) were isolated to construct a highly efficient consortium named (C4C5 + L3L4). When the compound bacteria agent combination by VC4: VC5: VL3: VL4 = 1:2:2:1, the crude oil degradation efficiency of 7 days was stable at 50.63% ~ 55.43% under different conditions. Degradation mechanism was analyzed by FTIR, GC-MS and IC technology and the following conclusions showed that in the system of adding consortium (C4C5 + L3L4), the heavy components were converted into saturated and unsaturated components. The bacterial consortium could first degrade medium and long chain alkanes into short chain hydrocarbons and then further degrade. And the dechlorination efficiency of 2,4-DCP in the degradation system reached 73.83%. The results suggested that the potential applicability and effectiveness of the selected bacteria consortium for the remediation of oil-contaminated water or soil with the existence of chlorophenol compound.

Effects of Electroacupuncture on Expression of PI3K/Akt/Foxo3a in Granulosa Cells from Women with Shen (Kidney) Deficiency Syndrome Undergoing in vitro Fertilization-Embryo Transfer / 中国结合医学杂志

OBJECTIVE@#To observe the effects of electroacupuncture (EA) on reproductive outcomes in women with Shen (Kidndy) deficiency syndrome after in vitro fertilization-embryo transfer (IVF-ET), and explore the underlying molecular mechanism.@*METHODS@#Sixty-six infertile patients with Shen deficiency syndrome undergoing IVF-ET were divided into EA or control groups according to a random table, 33 cases in each group. Before undergoing IVF, patients in the EA and control groups received EA therapy and placebo needle puncture, respectively, for 3 menstrual cycles. Shen deficiency syndrome scores were assessed. Other outcome measures included the number of retrieved oocytes and fertilization, high-quality embryo and clinical pregnancy rates. Follicular fluid was collected on the day of oocyte retrieval, and granulosa cell expression of phosphatidylinositide 3-kinases (PI3K), serine-threonine kinase (Akt) and forkhead box O3 (Foxo3a) mRNA were measured by reverse transcribed and quantitative real-time polymerase chain reaction.@*RESULTS@#Syndrome scores for pre- versus post-treatments decreased significantly (16.53±1.75 to 8.67±1.61) in the EA group (P<0.05), but showed no significant change in the control group (17.18±1.58 to 14.74±1.58). A significant difference in score change was found between the EA and control groups (P<0.05). High-quality embryo and clinical pregnancy rates were both increased in the EA group compared with the control group [69.15% (195/282) vs. 60.27% (176/292) and 66.67% (22/33) vs. 42.42% (14/33), respectively, P<0.05]. The fertilization rate was equivalent in EA and control groups. No difference was found in the number of retrieved oocytes between the two groups. Granulosa cell expression levels of PI3K and Akt mRNA were significantly increased in the EA group compared with the control group, while the expression of Foxo3a was reduced (all P<0.05).@*CONCLUSIONS@#For infertile patients with Shen deficiency syndrome undergoing IVF, EA for tonifying Shen as an adjunct treatment may alleviate clinical symptoms and improve the high-quality embryo rate. The EA-induced mechanism may involve regulation of PI3K/Akt/Foxo3a expression in granulosa cells to improve the developmental microenvironment of oocytes and inhibit granulosa cell apoptosis, possibly contributing to the improved clinical pregnancy rate (Registration No. ChiCTR 1800016217).

Analysis of factors related to oil-spilling and establishment of limit standards of Asparagi Radix / 中国中药杂志

Some samples of Asparagi Radix were collected from medical markets.Colors of Asparagi Radix were observed by human vision and recorded to judge whether samples were degenerative.Water content of Asparagi Radix was determined by a drying method.The chroma value and color difference were determined and calculated by a colorimeter.With the deepening of color,the L*value was decreased and a*and ΔE*values were increased.It showed that the results determined by colorimeter can replace the results of visual observation.An HPLC method was established and used to determine the contents of 5-hydroxymethylfurfural(5-HMF) in Asparagi Radix.The results showed the 5-HMF contents were from 0.002 255 to 0.049 14 mg·g-1 in some samples with yellowish-white or yellowish-brown color,significantly increased from 0.080 80 to 0.105 1 mg·g-1 in some samples with brown color,and up to 1.033 mg·g-1 in an oil-spilling sample with dark brown color.This result demonstrated that the 5-HMF contents were significantly increased by accompanied with the deepening of color.There were the significant negatively correlation between the 5-HMF content and the L*value(P<0.01) and positively correlation between the 5-HMF content and the a*or ΔE*value(P<0.01) by the spearman analysis.The oil-spilling and qualified samples were clustered into two alone categories by the cluster analysis.That the limited standards of the 5-HMF content is not higher than 0.02% by HPLC method and of the L*value is not less than 50 by colorimeter method were suggested for Asparagi Radix.It is firstly reported the multiple-factor analysis about oil-spilling and discoloration and the establishment of limited standard of Asparagi Radix.

Effect of Yangjing Zhongyu Tang in Treating Kidney-Yin Deficiency Due to Repeated Implantation Failure and on LIF,VEGF,IL-1β,MCP-1 Levels / 中国实验方剂学杂志

Objective:To explore the clinical efficacy of Yangjing Zhongyu Tang in treating kidney-Yin deficiency due to repeated implantation failure (RIF). Method:Totally 70 patients with kidney yin deficiency due to repeated implantation failure (RIF) who received the periodic Frozen thawed embyo (FET) after in vitro fertilization (IVF) embryo cryopreservation were selected from the Integrative Medicine Reproductive and Genetics Center of Affiliated Hospital of Shandong University of Traditional Chinese Medicine(TCM). They were randomly divided into observation group and control group, with 35 cases in each group. All patients were received long protocol controlled ovarian hyperstimulation. Observation group was given Yangjing Zhongyu Tang from the 5th day to the 14th day of the menstrual cycle 1 month before transplantation, and continued to take it on the 5th day of the menstrual cycle in the month of the transplantation. Control group did not given TCM. After treatment, TCM syndrome scores, endometrial thickness, typing, uterine artery pulsation index (PI) and resistance index (RI) of HCG, endometrial secretion leukemia inhibitory factor (LIF), blood vessels endothelial growth factor (VEGF), interleukin-1β (IL-1β) and monocyte chemoattractant protein-1 (MCP-1), number of eggs obtained, fertilization rate and embryo transfer results were observed. Result:After treatment, the scores of TCM syndromes in observation group were significantly improved, with statistically significant difference (PPPβ and LIF of observation group were higher than those of control group, and the MCP-1 was lower than that of control group (PPConclusion:Yangjing Zhongyu Tang can significantly alleviate the clinical symptoms of patients with kidney-Yin deficiency due to RIF, effectively improve the type A endometrial morphology of RIF patients, reduce the uterine artery blood flow resistance, optimize endometrial implantation, and significantly improve the clinical pregnancy rate.

Monthly Dynamics of Physical and Chemical Indexes of Achyranthes Bidentatae Radix Stored in Simple and Cool Warehouses / 中国药学杂志

OBJECTIVE: To study the monthly dynamics of physical and chemical indexes in Achyranthis Bidentatae Radix(ABR, the dried root of Achyranthes bidentata Bl.) stored in simple and cool warehouses. METHODS: ABR was stored in simple and cool warehouses for 27 months. The color was observed. The water content was determined based on the drying method. The contents of β-ecdysone and 5-hydroxymethylfurfural (5-HMF) were determined by HPLC method. The accumulation of temperature difference between the simple and cool warehouses was evaluated with a relative temperature cumulation (RTC) method. The monthly dynamics of physical and chemical indexes of ABR was analyzed with RTC. RESULTS: As the extension of storage time, the ABR stored in the simple warehouse showed deeper color and harder texture, but the ABR stored in the cool warehouse still had soft texture without significant color change. The contents of β-ecdysone in ABR stored in the two warehouses both gradually decreased and dropped to lower than the limit of 0.030% ruled by China Pharmacopoeia when being stored for up to 27 months. The contents of 5-HMF of ABR stored in the two warehouses both increased and were higher for the sample in the simple warehouse than that in the cool warehouse. CONCLUSION: The concept of RTC is put forward and used to study the monthly dynamics of chemical constituents in traditional Chinese medicine during storage for the first time. The physical and chemical indexes of ABR varies during storage. Two years of storage time of ABR is suggested.

Identification of Ilicis Rotundae Cortex and Its Adulterant Based on HPLC Fingerprint / 中国中医药信息杂志

Objective To establish HPLC fingerprints for the identification of Ilicis Rotundae Cortex (Jiubiying), dried bark of Ilex rotunda Thunb., and its adulterant, dried bark of Ilex godajam (Colebr.) Wall.. Methods Some samples named Jiubiying were collected from some medicine markets and drugstores. The HPLC fingerprints were obtained to detect Jiubiying and Ilex godajam. The software Similarity Evaluation System of Chromatographic Fingerprints of Traditional Chinese Medicine (2004A edition) was used to evaluate fingerprints. Results The fingerprints of Jiubiying and Ilex godajam were established. Methodological study met the technical requirements of fingerprints. There were 31 and 28 common peaks in Jiubiying and Ilex godajam, respectively. Jiubiying and Ilex godajam could be classified into two clusters by principal component analysis and hierarchical clustering analysis. Conclusion Jiubiying and Ilex godajam can be identified by HPLC fingerprints. This method can provide an effective method for quality evaluation of Jiubiying.

Chemical properties and antioxidant activity of a water-soluble polysaccharide from Dendrobium officinale.

In this report, a water-soluble polysaccharide was obtained from the dried stems of Dendrobium officinale Kimura et Migo by hot-water (70-75°C) extraction and 85% ethanol precipitation, and successively purification by DEAE-cellulose anion-exchange chromatography and gel-permeation chromatography. The D. officinale polysaccharide (DOP) has a molecular weight of 8500Da. Monosaccharide composition analysis reveals that DOP is composed of mannose, glucose, and arabinose with a trace of galacturonic acid in a molar ratio of 6.2:2.3:2.1:0.1. Periodate oxidation-smith degradation and 1D and 2D NMR spectroscopy analysis suggest the predominance of mannose and glucose, and it contains a 2-O-acetylglucomannan and (1→4)-linked-ß-d-mannopyranosyl and (1→4)-linked-ß-d-glucopyranosyl residues. Atomic force microscope shows that DOP mainly exists as rod-shaped chains, supporting high degrees of polymerization. The antioxidant activities of the polysaccharide in vitro assay indicate that DOP has good scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, higher scavenging activity of hydroxyl radical, and metal chelating activities.

Effect of yougui formula granule on ovarian granulosa cells gene expression profiles in IVF patients of shen yang deficiency syndrome / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Yougui Formula Granule (YFG) on ovarian granulosa cells gene expression profiles in in vitro fertilization-embryo transfer (IVF-ET) patients of Shen yang deficiency syndrome (SYDS) from the viewpoint of genomics.</p><p><b>METHODS</b>Totally 72 infertility patients undergoing IVF-ET were randomly assigned to the treatment group and the control group according to random digit table, 36 in each group. Patients in the treatment group took YFG combined gonadotropin (Gn), while those in the control group took placebos combined Gn. All medication lasted for 3 menstrual cycles before IVF. With high-throughput gene sequencing technology, gene expression profiles of ovarian granulosa cells in the two groups were analyzed to explore the difference by gene ontology (GO) enrichment analysis and kyoto encyclopedia of genes and genomes (KEGG) pathway analysis.</p><p><b>RESULTS</b>Ovarian granulosa cell gene expression profiles from the follicular fluid showed, when compared with the control group, 391 differential genes were found in the syndrome-control group, 153 down-regulated and 238 upregulated. Enrichment of differentially expressed cellular location and molecular function of genes involved cell proliferation and apoptosis associated cyclin, protein ubiquitination, construction of microtubules and microfilament, mitochondrial function and energy-related factors, regulatory factors for hormone synthesis. Participated pathways involved energy metabolism pathway and transforming growth factor-β (TGF-β) signaling pathway.</p><p><b>CONCLUSIONS</b>There existed significant difference in gene expression profiles of ovarian granulosa cells between the treatment group and the control group. Differentially expressed genes involved in biological processes correlates with Shen yang deficiency induced proliferation of germ cells, confused apoptosis, and hindered process during which mitochondria produced energy.</p>

Effects of Liuwei Dihuang Granule (symbols; see text) on the outcomes of in vitro fertilization pre-embryo transfer in infertility women with Kidney-yin deficiency syndrome and the proteome expressions in the follicular fluid / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effects of Liuwei Dihuang Granule ([symbols; see text], LDG) for tonifying Kidney (Shen) on the outcomes of in vitro fertilization pre-embryo transfer (IVF-ET) of infertility women with Kidney-yin deficiency syndrome and to explore its mechanism by detecting the proteome expression in the follicular fluid.</p><p><b>METHODS</b>Sixty-six infertility patients of Kidney-yin deficiency syndrome who would undergo IVF-ET, were randomly assigned to a treatment group and a control group according to a random number table, 33 cases in each group. Another 33 cases of non-Kidney-yin deficiency syndrome was taken as a syndrome-control group. Besides Western routine therapy, LDG was given 3 menstrual cycles before IVF to the treatment group, and a placebo granule to the control and syndrome-control groups. The scores of Kidney-yin deficiency symptoms (sore waist and knees, dry vagina, dysphoria with feverish sensation in the chest, palms and soles, etc.) were assessed, the number of retrieved oocytes, rates of high quality oocytes and embryos, fertility rate and clinical pregnancy rate were recorded, and the follicular fluid was collected on the day when the ovum was picked up, the differential protein expression was detected using two-dimensional gel electrophoresis, and then, matrix assisted laser desorption ionization time-of flight mass spectrometry (MALDI-TOF-MS) was applied to identify the proteins.</p><p><b>RESULTS</b>The syndrome score in the treatment group decreased significantly from 16.09±2.58 to 8.67±2.13, while it changed insignificantly in the control group, with a significant difference in the lowering score between the two groups (P<0.05); the high quality rates of oocytes and embryos and clinical pregnancy rate were all superior in the treatment group to the control group (82.29% vs 78.08%, 76.76% vs 68.79%, 63.64% vs 36.36%, all P<0.05). The protein expression map from the follicular fluid showed that compared with the control group, 33 differential protein expressions were found in the syndrome-control group, among which 18 were down-regulated, and 15 up-regulated; in the treatment group 28 differential protein expressions were found, among which 15 were down-regulated, and 13 up-regulated. Through MALDI-TOF-MS, 14 proteins were identified (P<0.05).</p><p><b>CONCLUSIONS</b>For the infertility patients undergoing IVF, LDG could alleviate clinical symptoms, improve rates of high quality oocytes and embryos, so as to raise clinical pregnancy rate. The mechanism may be through regulating proteome expression in the follicular fluid to improve the developmental microenvironment for oocytes which would lead to a successful embryo implantation.</p>

Novel function of N,N-bis(2-chloroethyl)docos-13-enamide for reversal of multidrug resistance in tongue cancer.

Multidrug resistance (MDR) is a key element in the failure of chemotherapies, and development of agents to overcome MDR is crucial to improving cancer treatments. The overexpression of glutathione-S-transferases (GSTs) is one of the major mechanisms of MDR. Because some agents used in traditional Chinese medicine have strong antitumor effects coupled with low toxicity; we investigated the ability of N,N-bis(2-chloroethyl)docos-13-enamide (compound J), the synthesized analog of a highly unsaturated fatty acid from Isatis tinctoria L., to reverse the MDR induced by adriamycin (ADM) in TCA8113/ADM cells. We found that compound J significantly increased the cytotoxicity of ADM in TCA8113/ADM cells, with a reversal fold of 2.461. Analysis of the mechanisms through which compound J reversed MDR indicated that compound J significantly decreased the activity of GSTs and enhanced the depletion of GSH in TCA8113/ADM cells, but did not affect the P-glycoprotein (P-gp) efflux. Taken together, our data suggested that compound J was an excellent candidate for reversing MDR in cancer therapy.

[Determination of degree of esterification in pectic polysaccharides from Angelica sinensis (Oliv.) diels by Fourier transform infrared spectroscopy].

An infrared spectroscopy method for rapid quantitative determination of the degree of esterification (DE) of pectic polysaccharides was developed. Taking pectic polysaccharides from angelica sinensis as samples, the calibration curve was established between DE and the ratio of A1 730/(A1 730 + A1 630). The square of the linear correlation coefficient was 0.822, indicating a good linear relationship between the DE and the absorbent area at 1 730 and 1 630 cm(-1). Using this method, the DE of pectic polysaccharides from angelica sinensis was determined to be between 42.36% and 54.06% for crude and purified samples respectively. The relationship between infrared spectroscopy data and titrimetrically determined DE values was investigated, indicating an excellent reproducibility. This method is characterized by less sample, simple manipulation, higher sensibility, and speedy analysis compared to chemical method and other instrumentations. FTIR shows a good feasibility and can be a rapid, alternative method to titrimetric analysis for DE determination.

[Orthogonal experiment of the extraction of triterpenoids from stems of Hyptis suaveolens with microwave].

OBJECTIVE: To study the best technology of the extraction of triterpenoids from the stems of Hyptis suaveolens with microwave. METHODS: Orthogonal experiment was carried out to investigate 4 influential factors as follows: the time (A), the temperature (B), the solid fluid compared to (C), the NaOH density (D). RESULTS: The optimal conditions for microwave extraction were A1 B2 C3 D2. CONCLUSION: The microwave extraction can extract more triterpenoids from the stems of Hyptis suaveolens in shorter time with less energy. It also shows a promising prospect for leaching the effective constituents from Chinese herbal medicine by using microwave extraction.