Blockage of Orai1-Nucleolin interaction meditated calcium influx attenuates breast cancer cells growth.

As an important second messenger, calcium (Ca2+) regulates a wide variety of physiological processes. Disturbance of intracellular calcium homeostasis implicated in the occurrence of multiple types of diseases. Orai1 is the major player in mediating store-operated calcium entry (SOCE) and regulates calcium homeostasis in non-excitable cells. Over-expression and activation of Orai1 have been reported in breast cancer. However, its molecular mechanisms are still not very clear. Here, we demonstrated that Nucleolin (NCL) was a novel interacting partner of Orai1. NCL is a multifunctional nucleocytoplasmic protein and is upregulated in human breast tumors. The binding of C-termini of NCL (NCL-CT) to N-termini of Orai1 (Orai1-NT) is critical for mediating calcium influx and proliferation of breast cancer cells. Blocking the NCL-Orai1 interaction by synthesized Orai1 peptide can effectively reduce the intracellular calcium influx and suppress the proliferation of breast cancer cells in vitro and in vivo. Our findings reveal a novel activation mechanism of Orai1 via direct interaction with NCL, which may lead to calcium homeostasis imbalance and promote the proliferation of breast cancer cells. Blocking NCL-Orai1 interaction might be an effective treatment of breast cancer.

[Professor YIN Ke-jing's experience of Jingluo Bietong acupuncture technique for shoulder arthralgia].

Professor YIN Ke-jing's experience of Jingluo Bietong acupuncture technique for shoulder arthralgia is summarized. According to the location of pain, shoulder arthralgia is divided into hand-taiyin type, hand-yangming type, hand-shaoyang type, hand-taiyang type and mixed type. The acupoint selection is based on Jingluo Bietong acupuncture technique, and Shugu (BL 65) or Feiyang (BL 58) are selected for hand-taiyin type, Taichong (LR 3) for hand-yangming type, Taixi (KI 3) and Dazhong (KI 4) for hand-shaoyang type, and Taibai (SP 3) or Diji (SP 8) for hand-taiyang type. In addition, combined with acupuncture at the blocking points of meridians and Xiangu (ST 43), better efficacy could be obtained.

[Professor YIN Ke-jing's experience of treating pain syndrome from meridians and collaterals].

Professor YIN Ke-jing is good at identifying the location of the disease by using meridian examination. With rich knowledge of the classics, combined with his own clinical experience, professor YIN puts forward "spasmodic muscle-tendon leads to pain", and the effect of Guanci (acupuncture at joints) is superior. Moreover, according to Inner Canon of Yellow Emperor, he has summarized new method of jingluo bietong, which uses fewer acupoints but achieves satisfactory efficacy. This method has broadened the principles of acupoint selection and prescriptions for clinical treatment. This method for treating pain can harmonize qi-blood and smooth meridians, and improve the clinical effect.

Orlistat-Induced Gut Microbiota Modification in Obese Mice.

INTRODUCTION: Accumulating evidence has indicated that alterations of gut microbiota have been involved in various metabolic diseases. Orlistat, a reversible inhibitor of pancreatic and gastric lipase, has beneficial effects on weight loss and metabolism. However, the effect of orlistat on the composition of gut microbiota remains unclear. OBJECTIVE: We aimed to explore the effect of orlistat on gut microbiota in high-fat diet (HFD) fed C57BL/6J obese mice. METHODS: C57BL/6J mice were randomly divided into three groups: control (NCD), HFD, and HFD + orlistat (ORL). Mice in the NCD group were fed chow diet, while the other groups were fed HFD for 6 months, and orlistat was added in the final 3 months in the HFD + ORL group. After sacrifice, body weight and metabolic parameters were assessed, and the gut microbial composition was analyzed by 16S rRNA gene sequencing. RESULTS: Orlistat treatment exerted beneficial effects on body weight, plasma cholesterol, and glucose tolerance. Meanwhile, orlistat treatment modified the gut microbiota, presenting as reduced total microbial abundance and obvious upregulated bacteria. Moreover, the upregulated bacteria correlated with several metabolic pathways. CONCLUSIONS: Orlistat may exert beneficial effects on body weight and glucose tolerance through modifying the composition of gut microbiota.

Electrochemical biosensor based on Se-doped MWCNTs-graphene and Y-shaped DNA-aided target-triggered amplification strategy.

A highly sensitive electrochemical biosensor for detection of platelet-derived growth factor-BB (PDGF-BB) is developed by using Se-doped multi-walled carbon nanotubes (MWCNTs)-graphene hybrids as electrode supporting substrate, hemin/G-quadruplex as trace labels and Y-shaped DNA-aided target recycling as signal magnifier. The aptamer-containing hairpin probes were first immobilized on the electrode. When target PDGF-BB was added, the aptamer binded PDGF-BB to trigger catalytic assembly of two other hairpins to form many G-quadruplex Y-junction DNA structures, which released PDGF-BB to again bind the intact aptamer to initiate another assembly cycle. G-quadruplex/hemin complexes were produced when hemin was added to generate substantially amplified current output. The developed assay showed a linear range toward PDGF-BB from 0.1 pM to 10 nM with a detection limit of 27 fM (S/N = 3). The method showed excellent specificity and repeatability, and could be expediently applied for sensitive detection of other molecules by simply changing the aptamers.

Relationship between total polar components and polycyclic aromatic hydrocarbons in fried edible oil.

Deep-fried dough sticks (a Chinese traditional breakfast) were fried individually in peanut, sunflower, rapeseed, rice bran, soybean and palm oil without any time lag for 32 h (64 batches fried, each for 30 min) and fried oil samples were obtained every 2 h. The frying-induced changes in the levels of total polar compounds (TPC) and polycyclic aromatic hydrocarbons (PAHs) were investigated by edible oil polar compounds (EOPC) fast separation chromatographic system and gas chromatography-mass spectrometry (GC-MS), respectively. The correlations were analysed of TPC with benzo[a]pyrene (BaP), TPC and PAH4 (benzo[a]anthracene, chrysene, benzo[b]fluoranthene and benzo[a]pyrene) as well as TPC with PAH16 (USEPA 16 PAHs). The results revealed that the levels of TPC and PAHs in fried oil considerably increased with frying time, and the type of oil affected their formation, which could inform the choice of oil for frying. The total BaP equivalents (∑BaPeq) concentrations in fresh oil and in oil whose TPC exceeded 27% were 2.14-13.48 and 5.78-10.80 µg kg-1, respectively, which means that the carcinogenic potency of frying oil was more pronounced than that of fresh oil. In addition, the TPC concentration was significantly correlated with the concentrations of the sum of the 16 PAHs, PAH4 and BaP, so that the levels of PAHs could be predicted according to the levels of TPC in fried oil. In European standards, the rejection point for TPC in frying oil should be recalculated when considered PAHs. In all, the concentration of PAHs is a vital factor for ensuring the safety of frying oil.

BODIPY-Mn nanoassemblies for accurate MRI and phototherapy of hypoxic cancer.

Hypoxia promotes not only the metastasis of tumors but also therapeutic resistance. Photosensitizer-mediated consumption of O2 during photodynamic therapy (PDT) reinforces tumor hypoxia. Herein, a light-dependent attenuator of a hypoxic environment is reported for accurate MRI and phototherapy of hypoxic cancer. First, a photoresponsive Mn(ii) nanoassembly was constructed, then it was assembled with bovine serum albumin (BSA) and modified with polyethylene glycol-folic acid (PEG-FA), forming cancer targeting Mn-DBA@BSA-FA nanoassemblies, which offer T1 signals and can catalyze the water oxidation reaction under irradiation of red light emitting diode (LED) light with the generation of O2 and heat. Moreover, they could selectively penetrate through and accumulate in the tumor tissues with clear T1 magnetic resonance imaging (MRI) signals, and have remarkably eliminated the tumors in vivo, while they are of low toxicity to the healthy organs. The release of the Mn(ii) complex from the nanoparticles in an acidic environment and the in vivo biodistribution results confirm the selective cancer targeting. Our work demonstrates the potential of nanoparticles as excellent theranostic agents for MR imaging combined with phototherapy triggered by near-infrared light.

Liraglutide restores angiogenesis in palmitate-impaired human endothelial cells through PI3K/Akt-Foxo1-GTPCH1 pathway.

Glucagon-like peptide-1 (GLP-1) and its analogues have a beneficial role in cardiovascular system. Here, we aimed to investigate whether liraglutide, a GLP-1 analogue, modulated angiogenesis impaired by palmitic acid (PA) in cultured human umbilical vein endothelial cells (HUVECs). Cells were incubated with liraglutide (3-100 nmol/L) in the presence of PA (0.5mmol/L), and endothelial tube formation was observed and quantified. The protein levels of signaling molecules were analyzed and the specific inhibitors were used to identify the signaling pathways through which liraglutide affected angiogenesis. Results showed that liraglutide ameliorated endothelial tube formation impaired by PA in HUVECs in a dose-dependent manner. Meanwhile, liraglutide increased the phosphorylation of Akt and forkhead box O1 (Foxo1), and upregulated the levels of guanosine 5'-triphosphate cyclohydrolase 1 (GTPCH1) and endothelial nitric oxide synthase (eNOS) in PA-impaired HUVECs. Notably, addition of the PI3K inhibitor LY294002, Foxo1 nuclear export inhibitor trifluoperazine dihydrochloride (TFP), GTPCH1 inhibitor 2,4-diamino-6-hydroxypyrimidine (DAHP) or NOS inhibitor N-nitro-l-arginine-methyl ester (L-NAME) eliminated the angiogenic effect of liraglutide. Moreover, either LY294002 or TFP abolished the liraglutide-induced upregulation of GTPCH1 and eNOS protein levels. In conclusion, liraglutide restores angiogenesis in PA-impaired HUVECs. The effect is mediated via upregulation of GTPCH1 and eNOS levels in a PI3K/Akt-Foxo1-dependent mechanism.

Ultrasensitive sensing platform for platelet-derived growth factor BB detection based on layered molybdenum selenide-graphene composites and Exonuclease III assisted signal amplification.

A highly sensitive and ultrasensitive electrochemical aptasensor for platelet-derived growth factor BB (PDGF-BB) detection is fabricated based on layered molybdenum selenide-graphene (MoSe2-Gr) composites and Exonuclease III (Exo III)-aided signal amplification. MoSe2-Gr is prepared by a simple hydrothermal method and used as a promising sensing platform. Exo III has a specifical exo-deoxyribonuclease activity for duplex DNAs in the direction from 3' to 5' terminus, however its activity is limited on the duplex DNAs with more than 4 mismatched terminal bases at 3' ends. Herein, aptamer and complementary DNA (cDNA) sequences are designed with four thymine bases on 3' ends. In the presence of target protein, the aptamer associates with it and facilitates the formation of duplex DNA between cDNA and signal DNA. The duplex DNA then is digested by Exo III and releases cDNA, which hybridizes with signal DNA to perform a new cleavage process. Nevertheless, in the absence of target protein, the aptamer hybridizes with cDNA will inhibit the Exo III-assisted nucleotides cleavage. The signal DNA then hybridizes with capture DNA on the electrode. Subsequently, horse radish peroxidase is fixed on electrode by avidin-biotin reaction and then catalyzes hydrogen peroxide and hydroquinone to produce electrochemical response. Therefore, a bridge can be established between the concentration of target protein and the degree of the attenuation of the obtained signal, providing a quantitative measure of target protein with a broad detection range of 0.0001-1 nM and a detection limit of 20 fM.

Attitudes toward the American nutrition guidelines for the critically ill patients of Chinese intensive care physicians.

Nutrition therapy is essential for the management of critically ill patients. Some guidelines have been published to standardize and optimize the nutrition therapy. However, there are still many controversies in nutrition practice and there is a gap between guidelines and clinical nutrition therapy for patients in intensive care units (ICUs). This study aimed to assess attitudes and beliefs toward nutrition therapy of Chinese intensive care physicians by using the American guidelines as a surrogate. A questionnaire was sent to 45 adult ICUs in China, in which surveyed physicians were asked to rate their attitudes toward the American guidelines. A total of 162 physicians from 45 ICUs returned the questionnaires. Physicians were categorized into groups according to their professional seniority, hospital levels and whether they were members of Chinese Society for Parenteral and Enteral Nutrition (CSPEN). Overall, 94% of the respondents thought that nutrition therapy for critically ill patients was very important, and 80% mentioned that they used the American guidelines. There was diversity of opinion on the recommendations pertaining to nutrition assessment, supplemental parenteral nutrition and cutoff values for gastric residual volume, negative or neutral attitudes about these recommendations were 43%, 59% and 41%, respectively. Members of CSPEN were more likely to select a greater strength of recommendation than non-members. In conclusion, the overall attitudes of Chinese intensive care physicians toward the American guidelines were positive. Nevertheless, given the great guideline-practice gap, nutrition-focused education is warranted for many intensive care physicians in China.

Ultrasensitive biosensing platform based on layered vanadium disulfide-graphene composites coupling with tetrahedron-structured DNA probes and exonuclease III assisted signal amplification.

Recently, two-dimensional layered transition metal dichalcogenide materials have been very fascinating in electrochemical fields due to their unique electronic and electrochemical properties. In the present work, a novel VS2-graphene (VS2-GR) composite is firstly prepared by a facile one-step hydrothermal method. A sensitive, universal and label-free electrochemical aptamer-based sensor (aptasensor) is then designed to detect the platelet-derived growth factor BB (PDGF-BB) based on VS2-GR coupled with Exo III-aided autocatalytic signal amplification. In the absence of PDGE-BB, the aptamer hybridized with the complementary DNA (cDNA) and the single-strand signal DNA labeled with biotin at 5' end cannot be cleaved by the Exo III. When the above mixture is applied on the tetrahedron-structured probe (T-DNA) modified AuNP/VS2-GR electrode, signal DNA hybridizes with the protruding of T-DNA and a large number of avidin-HRP are adsorbed on the modified electrode by the specific binding of avidin-biotin, which leads to a strong current response by the catalysis of HRP to the mixture of hydrogen peroxide and hydroquinone (H2O2 + HQ). However, in the presence of PDGF-BB, the aptamer is preferentially associated with PDGF-BB, which facilitates the formation of double-strand DNA between cDNA and signal DNA. The duplex DNA is digested by Exo III from the 3' blunt terminus of signal DNA and the cDNA is released. The liberative cDNA then reacts with the remaining signal DNA in the solution to perform a new cleavage process. Finally, the cyclic hybridization-hydrolysis process leads to a great decrease of biotin on the electrode, thus reducing the quantity of signal molecule HRP and causing an obvious decline of current response. This proposed protocol exhibits high specificity, good sensitivity, and good stability for PDGF-BB detection. Moreover, the aptasensor can be applied as a universal assay for the detection of various targets including proteins, DNAs, and metal ions.

The investigation of the interactions between CdSe quantum dots and human serum albumin by resonance Rayleigh scattering and second-order scattering spectra.

Human serum albumin (HSA) was the most abundant protein in human plasma and has significant physiological function. In Tris-HCl buffer solution (pH 7.4), water-soluble semiconductor CdSe quantum dots (QDs) reacted with HSA and the products resulted in a great enhancement of the intensity of resonance Rayleigh scattering (RRS) and second-order scattering (SOS). Based on this, a new method was developed to investigate the interactions between QDs and HSA. The parameters with regard to determination were optimized, and the reaction mechanism was discussed. Under optimal conditions, the increments of scattering intensity (DeltaI) were directly proportional to the concentrations of HSA in the range of 0.4-48.0 micromol L(-1). The detection limits were 0.10 micromol L(-1) for RRS method and 0.25 micromol L(-1) for SOS method. The proposed method was sensitive, simple and rapid. It has been successfully applied to the determination of HSA in human urine samples. Analytical results obtained with this novel assay were satisfactory.

Therapeutic effect of compound danshen solution on hemorrhagic shock combined with coagulopathy in rats / 中国实验血液学杂志

To investigate the effects of complex danshen solution and heparin on the changes of blood coagulation factors in rats with hemorrhagic shock, and to explore the therapy of coagulopathy by compound danshen solution, the rat model of hemorrhagic shock was set up, 40 SD rats were randomized into four groups: sham operation, shock, compound danshen solution and heparin groups, each group was composed of 10 SD rats. Plasma SFMC, TM, ATIII, D-D, t-PA, PAI levels and APTT were detected, incidences of bleeding complications between heparin and danshen group were compared. The results showed that plasma SFMC, D-D levels in shock group were higher but ATIII level in shock group was lower than that in sham operation group, compound danshen solution group and heparin group (P < 0.001), TM levels obviously increased in shock group and heparin group (P < 0.001). There was no significant difference between compound danshen solution and sham-operation groups. Plasma t-PA, D-D levels obviously increased after shock for 2 hours, PAI level reached the peak after shock for 4 hours, but t-PA decreased. After shock for 6 hours, plasma PAI descended, t-PA continually drop in, but PAI and D-D remained in higher levels. Plasma D-D level in heparin group was lower than that in shock group, t-PA level was higher than that in shock group, but there was no significant difference between in heparin and shock groups. Plasma t-PA, PAI and D-D levels in compound danshen solution group were lower than that in shock group. APTT of danshen group was lower than that of shock group and heparin group. Bleeding incidences was 30% in heparin group and 0% in danshen group, respectively. It is concluded that compound danshen solution may used to treat hypercoagulation and hyperfibrinolysis. In comparsion with heparin, danshen posses-ses advantages of safety with less bleeding complication and needs not tight monitor.

[Clinical study on effect of zhifeng huatan pingxiao recipe in treating attack stage of child asthma].

OBJECTIVE: To observe the therapeutic effect of Zhifeng Huatan Pingxiao recipe (ZHPR) in treating attack stage of child asthma. METHODS: Sixty child asthma patients with attack of mild and moderate degree were randomly divided into the treated group (40 patients treated with ZHPR) and the control group (20 patients treated with western medicine). The therapeutic effect and laboratory indexes, including peripheral eosinophil count (EOS), serum immunoglobulin E (IgE) and plasma endothelin (ET), before and after treatment were observed. RESULTS: The total effective rate in the treated group was 72.5%, which was not different to that in the control group (P > 0.05). The therapeutic effect between different TCM types (heat and cold types) in the treated group was not different significantly (P > 0.05). ET, IgE and EOS levels all lowered after treatment in the treated group, but as compared with those in the control group, the difference was insignificant (P > 0.05). CONCLUSION: The therapeutic effect of ZHPR in treating child asthma attack stage is equivalent to that of western medicine, it shows actions in improving laboratory indexes.

Clinical study on effect of zhifeng huatan pingxiao recipe in treating attack stage of child asthma / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effect of Zhifeng Huatan Pingxiao recipe (ZHPR) in treating attack stage of child asthma.</p><p><b>METHODS</b>Sixty child asthma patients with attack of mild and moderate degree were randomly divided into the treated group (40 patients treated with ZHPR) and the control group (20 patients treated with western medicine). The therapeutic effect and laboratory indexes, including peripheral eosinophil count (EOS), serum immunoglobulin E (IgE) and plasma endothelin (ET), before and after treatment were observed.</p><p><b>RESULTS</b>The total effective rate in the treated group was 72.5%, which was not different to that in the control group (P > 0.05). The therapeutic effect between different TCM types (heat and cold types) in the treated group was not different significantly (P > 0.05). ET, IgE and EOS levels all lowered after treatment in the treated group, but as compared with those in the control group, the difference was insignificant (P > 0.05).</p><p><b>CONCLUSION</b>The therapeutic effect of ZHPR in treating child asthma attack stage is equivalent to that of western medicine, it shows actions in improving laboratory indexes.</p>