[Study on quantification method of non-starch polysaccharides in Dendrobium catenatum].

The non-starch polysaccharides,mainly composed of glucomannans,are the major bioactive compounds in Dendrobium catenatum. In order to evaluate the quality of the medicinal materials and guide the production and processing,a quantification method of non-starch polysaccharides was established by stems of D. catenatum C15 strain collected from the pear epiphytic cultivation. The non-starch polysaccharides were obtained by " water extraction,α-amylase pretreatment,and alcohol precipitation once" method. The contents of starches,non-starch polysaccharides and monosaccharides were analyzed. In addition,the system suitability was tested. Compared with method of the Chinese Pharmacopoeia( 2015 edition),the contents of total polysaccharides,glucose,and mannose were decreased by 20. 9%,58. 8% and 1. 6% respectively. The method effectively digested starch and retained non-starch polysaccharides,and the analysis result was accurate and repeatable. Therefore,it is suitable for the content measurement of non-starch polysaccharides of D. catenatum. Furthermore,it could be an alternative method for quality control of D. catenatum and a reference in the determination of non-starch polysaccharides in other starch-containing medicinal materials.

Actuation of adaptive liquid microlens droplet in microfluidic devices: A review.

A significant growth of research on adaptive liquid lens is achieved over the past decades, and the field is still attracting increasing attentions, focusing on the transition from the current stage to the commercialized stage. The challenges faced are not limited to fabrication, material, small tuning range in focal lengths, additional control systems, limitations in special actuation methods and so on. In addition, the use of external driving parts or systems induce extra problem on bulky appearance, high cost, low reliability etc. Therefore, adaptive liquid lens will be an interesting research focus in both microfluidics and optofluidics science. This review attempts to summarize and focus on the droplet profile deformation under different driving mechanisms in tunable liquid microlens as well as the application in cameras, cell phone and so on. The driving techniques are generally categorized in terms of mechanisms and driving sources.

Droplet Manipulation under a Magnetic Field: A Review.

The magnetic manipulation of droplets is one of the emerging magnetofluidic technologies that integrate multiple disciplines, such as electromagnetics, fluid mechanics and so on. The directly driven droplets are mainly composed of ferrofluid or liquid metal. This kind of magnetically induced droplet manipulation provides a remote, wireless and programmable approach beneficial for research and engineering applications, such as drug synthesis, biochemistry, sample preparation in life sciences, biomedicine, tissue engineering, etc. Based on the significant growth in the study of magneto droplet handling achieved over the past decades, further and more profound explorations in this field gained impetus, raising concentrations on the construction of a comprehensive working mechanism and the commercialization of this technology. Current challenges faced are not limited to the design and fabrication of the magnetic field, the material, the acquisition of precise and stable droplet performance, other constraints in processing speed and so on. The rotational devices or systems could give rise to additional issues on bulky appearance, high cost, low reliability, etc. Various magnetically introduced droplet behaviors, such as deformation, displacement, rotation, levitation, splitting and fusion, are mainly introduced in this work, involving the basic theory, functions and working principles.

[Analysis of Microbial Interaction Law of Mud Membrane in IFAS Process for Treating Low Carbon Source Sewage in South China].

To assess the problem of sewage treatment under the condition of low carbon sources, we carried out a study of activated sludge and a biofilm symbiosis system (IFAS). The occurrence characteristics and interaction law of microorganisms in two phases of sludge membrane under low carbon source conditions were discussed, and their niche and influence on treatment efficiency were clarified. Through a pilot-scale experiment in actual water plants, the biofilm characteristics, sludge membrane activity, and succession law of flora were analyzed, and the microbial structure and interaction in sludge membrane in two phases under the control of different activated sludge ages were compared. The results showed that the sludge concentration in the reactor increased with the increase in SRT under variable SRT. Because the microbial concentration in SRT-H was much higher than that in SRT-L, the competition between mud films in SRT-H was more intense than that in SRT-L, and the pollutant removal efficiency in SRT-H was lower than that in SRT-L. Under the condition of low-carbon feed water, the sludge activity in the IFAS process decreased with the increase in SRT. Under the condition of low SRT(5 d), the nitrification, denitrification, phosphorus accumulation, and phosphorus absorption rate of activated sludge increased by 122%, 88%, 34%, and 44%, respectively, compared with that of high SRT (25 d). However, SRT had little effect on biofilm activity, and there was little difference in nitrification activity and denitrification activity between the two SRTs. Microbial sequencing analysis showed that the functional bacteria of the IFAS process were enriched and transferred with the change in SRT between the two phases of mud membrane. In SRT-L, the functional bacteria that were enriched and transferred between the two phases of mud film owing to the "seeding" effect were mainly unclassified_g__Enterobacteriaceae, whereas in SRT-H, Acinetobacter was mainly used. At the same time, by analyzing the distribution of dominant functional bacteria, it was found that there was some competition between denitrifying bacteria and phosphorus-accumulating bacteria in activated sludge. Under the condition of a lack of organic substrate in the influent, the relative abundance of denitrifying bacteria was obviously higher than that of phosphorus-accumulating bacteria, which indicated that denitrifying bacteria could better adapt to low-carbon source conditions. Thus, they could occupy a dominant competition position, which was mainly reflected in the increase in the relative abundance of aerobic denitrifying bacteria. In addition, the SRT change in the mud phase reacted in the membrane phase, making the residence time of biofilm change correspondingly, thus changing the flora structure, screening out different dominant bacteria genera, and further increasing the difference.

Anticancer effects of imperatorin isolated from Angelica dahurica: induction of apoptosis in HepG2 cells through both death-receptor- and mitochondria-mediated pathways.

BACKGROUND: Imperatorin (IM) is a furanocoumarin isolated from the root of Angelica dahurica, which is reported to have anticonvulsant and anticancer effects. In this study, the antiproliferative effect of IM on 9 human cancer cell lines was examined, and human hepatoma HepG2 cells were chosen as the target for preferential killing by IM. Particularly, the mechanism of IM-induced apoptosis and in vivo animal effects were also studied. METHODS: Cell viability was measured using MTT assay, and apoptosis was detected by Hoechst staining, annexin V-PI staining, and DNA laddering assay. Mitochondrial membrane potential was detected by JC-1 staining. Western blot analysis was employed to detect the expression of apoptosis-related proteins. In addition, the in vivo anticancer effect of IM was examined in nude mice bearing HepG2 cells. RESULTS: IM inhibited the proliferation of HepG2 cells through apoptosis induction in a time- and dose-dependent manner by observation of the nuclear morphology, DNA fragmentation, phosphatidylserine externalization, loss of mitochondrial membrane potential, release of cytochrome c into cytosol, and activation of caspase-3, caspase-8, caspase-9, and poly(ADP-ribose) polymerase cleavage. As cell death could partly be prevented by the caspase-8 or caspase-9 inhibitor and was evidenced by the results of Western blot analysis, our results also suggest that IM-induced apoptosis is mediated through both death receptor and mitochondrial pathways. In the animal model, IM was found to effectively suppress tumor growth by 31.93 and 63.18% at dosages of 50 and 100 mg/kg, respectively, after treatment for 14 days. No significant weight loss or toxicity to the hosts was found. CONCLUSIONS: IM can function as a cancer suppressor by inducing apoptosis in HepG2 cells through both death-receptor- and mitochondria-mediated pathways. Furthermore, the in vivo antitumor activities of IM are significant with negligible weight loss and damage to the host.

Expression and biochemical characterization of a multifunctional glycosidase from the thermophilic Bacillus licheniformis SR01.

A gene (gkdA) (741 bp) encoding a putative protein of 247 amino acids was cloned from the Bacillus licheniformis SR01. The protein was expressed in Escherichia coli BL21 with a molecular mass estimated by SDS-PAGE of approximately 28.03 kDa and showed a calculating isoelectric point (pI) of 6.42. Structure analysis and function identification showed that the enzyme was a multifunctional glycosidase. Its specific activity was 0.013 U/µg. The recombinant glycosidase showed a maximum activity at 50°C and pH 7.0. It was very stable below 90°C and may have heat activation at higher temperatures. The relative residual activity was still more than 80% after 120 min at pH 5.0-10.0. The enzyme activity was inhibited by Cu2+, Fe2+, Ca2+, Mg2+, Co2+, Li+, SDS and EDTA, activated by Ca2+, and not affected by Mn2+ and K+. Under simulated stomach, and in vitro intestine, conditions, the enzyme retained 80%, and more than 100%, activity, respectively, after incubation for 90 min. The excellent properties of this enzyme, specifically its thermal stability and multifunctional abilities, give it potential application in the field of feed processing and other high-temperature processing industries.

A sensitive and practical RP-HPLC-FLD for determination of the low neuroactive amino acid levels in body fluids and its application in depression.

Ion-exchange high performance liquid chromatography (HPLC) generally fails as a method to determine low levels of free amino acids (AAs) in body fluids. Here we present a modified reversed-phase HPLC (RP-HPLC) protocol for the determination of AAs in body fluids and its application in mood disorder patients. We improved a previous research protocol by modifying i) sample preparation, including deproteination, ii) derivitization, including derivating agent and condition, and iii) sample separation, which is mainly determined by the pH value, the components and the additives of the mobile phases. The combination of these modifications, together with fluorescence detection (FLD), allows sensitive and practical determination of free AA levels in body fluids of depressive patients. This protocol was validated by determining the postmortem cerebrospinal fluid (CSF) glutamic acid (Glu) and γ-aminobutyric acid (GABA) levels of 8 major depressive disorder (MDD) patients, 9 bipolar disorder (BD) patients, and 19 well-matched controls, while also testing the plasma and CSF AA levels of living MDD patients. CSF Glu and GABA levels were both significantly decreased in MDD but not in BD patients. The data indicate that this RP-HPLC-FLD protocol is applicable for detection of low levels of neuroactive AAs in body fluids, as well as for routine clinical applications.

Synthesis of arbutin by two-step reaction from glucose.

Arbutin was synthesized from glucose by two-step reaction below: (a) 2,3,4,6-tetra-O-acetyl-alpha-D-glucosyl chloride or bromide was prepared by glucose and acetyl halide (chloride or bromide). (b) 2,3,4,6-tetra-O-acetyl-alpha-D-glucosyl halide (Cl, Br) reacted with hydroquinone, methanol as solvent at pH=9.5-10.0.

Celastrol-induced apoptosis in human HaCaT keratinocytes involves the inhibition of NF-κB activity.

Psoriasis is a chronic inflammatory skin disease affecting 1-3% of the world's population. Traditional Chinese medicines have been extensively used for treating psoriasis with promising clinical results. Celastrol, a triterpenoid isolated from a Chinese herb Celastrus orbiculatus caulis, has been known to have diverse pharmacological effects such as anti-inflammatory, anti-cancer and antioxidant activities. The present study aimed at evaluating the anti-proliferative action of celastrol on cultured HaCaT cells and elucidating the mechanisms of action involved. Celastrol was shown to inhibit HaCaT cells growth with an IC50 value of 1.1 µM as measured by MTT assay. The ability of celastrol to induce apoptosis was studied by flow cytometric and western blot analyses. Celastrol was found to be capable of inducing apoptosis in HaCaT cells as characterized by phosphatidyl-serine (PS) externalization, depolarization of mitochondrial membrane potential and activation of caspase-3. The apoptosis induced by celastrol could be suppressed by Z-IETD-FMK and Z-LEHD-FMK, the respective caspase-8 and caspase-9 inhibitor. In addition, western blot analysis revealed a significant augmentation in the protein expression of Bax and attenuation in Bcl-2, suggesting that the celastrol-induced apoptosis acts through both death receptor and mitochondrial pathways. Moreover, western blot analysis on the expression of Rel/NF-κB demonstrated that the celastrol-mediated apoptosis on HaCaT cells was associated with the inhibition of the NF-κB pathway. Taken together, the present project has for the first time identified celastrol as a naturally occurring compound with potent apoptogenic action on cultured human keratinocytes, rendering it a promising candidate for further development into an anti-psoriatic agent.

Suillin from the mushroom Suillus placidus as potent apoptosis inducer in human hepatoma HepG2 cells.

During the search of new anti-cancer agent from high fungi, the ethyl acetate extract of the mushroom Suillus placidus was found to exhibit a significant cytotoxic activity against human hepatoma HepG2 cells. With bioassay-guided fractionation, a cytotoxic component suillin was isolated from the extract. The anti-cancer effect of suillin was subsequently examined in 8 human cancer cell lines by using MTT assay. It is of interest to note that human liver cancer cells (HepG2 cells, Hep3B cells, and SK-Hep-1) were preferentially killed by suillin with an IC(50) of approximately 2microM in a 48h treatment. Mechanistically. suillin was found for the first time to induce apoptosis in HepG2 cells as characterized by DNA fragmentation, phosphatidyl-serine (PS) externalization, activation of caspase-3, -8, -9, depolarization of mitochondrial membrane potential, as well as release of cytochrome c into the cytosol. Moreover, the apoptosis induced by suillin was suppressed by both caspase-8 and -9 inhibitors. Western blot analysis revealed significant increases in the protein levels of Fas death receptor, adaptor FADD protein, pro-apoptotic protein Bad and a decline of Bid. These results suggest that the induction of apoptosis by suillin is through both death receptor and mitochondrial pathways. Taken together, our results suggest that suillin might be an effective agent to treat liver cancer.

Macrocalyxin I.

The title compound, 2-[1,2,3,4,4a,4b,5,6,7,8,8a,9-dodecahydro-7-hydroxy-4b,8,8-trimethylphenanthren-2-yl]propenoic acid, C20H30O3, is a naturally occurring diterpenoid which was isolated from Rabdosia macrocalyx. The hydroxy and carboxy groups, which are located at the two ends of the molecule, both serve as simultaneous hydrogen-bond donors and acceptors. Two intermolecular O-H...O hydrogen bonds are present and link each molecule to four neighbours, thus forming an extensive hydrogen-bond network within the crystal.