Shexiang Tongxin Dropping Pill Promotes Angiogenesis through VEGF/eNOS Signaling Pathway on Diabetic Coronary Microcirculation Dysfunction.

OBJECTIVE: To study the effect of Shexiang Tongxin Dropping Pill (STDP) on angiogenesis in diabetic cardiomyopathy mice with coronary microcirculation dysfunction (CMD). METHODS: According to a random number table, 6 of 36 SPF male C57BL/6 mice were randomly selected as the control group, and the remaining 30 mice were injected with streptozotocin intraperitoneally to replicate the type 1 diabetes model. Mice successfully copied the diabetes model were randomly divided into the model group, STDP low-dose group [15 mg/(kg·d)], medium-dose group [30 mg/(kg·d)], high-dose group [60 mg/(kg·d)], and nicorandil group [15 mg/(kg·d)], 6 in each group. The drug was given by continuous gavage for 12 weeks. The cardiac function of mice in each group was detected at the end of the experiment, and coronary flow reserve (CFR) was detected by chest Doppler technique. Pathological changes of myocardium were observed by hematoxylin-eosin staining, collagen fiber deposition was detected by masson staining, the number of myocardial capillaries was detected by platelet endothelial cell adhesion molecule-1 staining, and the degree of myocardial hypertrophy was detected by wheat germ agglutinin staining. The expression of the vascular endothlial growth factor (VEGF)/endothelial nitric oxide synthase (eNOS) signaling pathway-related proteins in myocardial tissue was detected by Western blot. RESULTS: Compared with the model group, medium- and high-dose STDP significantly increased the left ventricular ejection fraction and left ventricular fraction shortening (P<0.01), obviously repaired the disordered cardiac muscle structure, reduced myocardial fibrosis, reduced myocardial cell area, increased capillary density, and increased CFR level (all P<0.01). Western blot showed that high-dose STDP could significantly increase the expression of VEGF and promote the phosphorylation of vascular endothelial growth factor receptor 2, phosphoinositide 3-kinase, protein kinase B, and eNOS (P<0.05 or P<0.01). CONCLUSION: STDP has a definite therapeutic effect on diabetic CMD, and its mechanism may be related to promoting angiogenesis through the VEGF/eNOS signaling pathway.

Improving efficiency and reducing enzyme inactivation during lipase-mediated epoxidation of α-pinene in a double-phase reaction system.

Chemoenzymatic epoxidation of olefin mediated by lipase is a green and environmentally friendly alternative process. However, the mass transfer barrier and lipase deactivation caused by the traditional organic-water biphasic reaction system have always been the focus of researchers' attention. To overcome these issues, we investigated the effects of reaction temperature and two important substrates (H2O2 and acyl donor) on the epoxidation reaction and interfacial mass transfer. As a result, we determined the optimal reaction conditions: a temperature of 30 °C, 30 wt-% H2O2 as the oxygen source, and 1 M lauric acid as the oxygen carrier. Additionally, by simulating the conditions of shaking flask reactions, we designed a batch reactor and added a metal mesh to effectively block the direct contact between high-concentration hydrogen peroxide and the enzyme. Under these optimal conditions, the epoxidation reaction was carried out for 5 h, and the product yield reached a maximum of 93.2%. Furthermore, after seven repetitive experiments, the lipase still maintained a relative activity of 51.2%.

IRF5 knockdown reverses TDP-related phenotypes partially by increasing TBK1 expression.

Interferon-regulatory factor 5 (IRF5) participates in the regulation of apoptosis, affects the phenotype of inflammatory macrophages and plays an essential role in the inflammatory response. However, the role of IRF5 in the progression of amyotrophic lateral sclerosis (ALS) remains largely unknown. Here, we show that IRF5 mainly accumulated in the nucleus in cells expressing the truncated 25 kD C-terminal fragments of TDP-43 (TDP-25, named TDP-25 cells hereafter). IRF5 knockdown using a lentivirus carrying an shRNA in TDP-25 cells exerted a protective effect and reduced the level of the apoptosis-related protein cleaved caspase-9 and the cell cycle arrest protein p21, while increasing the expression of the antioxidant transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) and its target molecule glutamate-cysteine ligase modulatory subunit (GCLM). Furthermore, IRF5-knockdown cells showed improved mitochondrial swelling and cristae dilation. In addition, we found that IRF5 mediated neuronal injury partly through the negative regulation of TANK-binding kinase 1 (TBK1). These data indicate that the loss of IRF5 in TDP-25 cells exerts a protective effect mainly by inhibiting apoptosis, regulating cell cycle arrest and alleviating oxidative stress.

Evaluation of the transverse aortic constriction model in ICR and C57BL/6J mice.

Transverse aortic constriction (TAC) is a frequently used model to investigate pressure overload-induced progressive heart failure (HF); however, there is considerable phenotypic variation among different mouse strains and even sub-strains. Moreover, less is known about the TAC model in ICR mice. Therefore, to determine the suitability of the ICR strain for TAC-induced HF research, we compared the effects of TAC on ICR and C57BL/6J mice at one, two and four weeks post-TAC via echocardiography, organ index, morphology, and histology. At the end of the study, behavior and gene expression patterns were assessed, and overall survival was monitored. Compared to the sham-operated mice, ICR and C57BL/6J mice displayed hypertrophic phenotypes with a significant increase in ventricle wall thickness, heart weight and ratio, and cross-sectional area of cardiomyocytes after a 2-week TAC exposure. In addition, ICR mice developed reduced systolic function and severe lung congestion 4 weeks post-TAC, whereas C57BL/6J did not. Besides, ICR mice demonstrated comparable survival, similar gene expression alteration but severer fibrotic remodeling and poor behavioral performance compared to the C57BL/6J mice. Our data demonstrated that ICR was quite sensitive to TAC-induced heart failure and can be an ideal research tool to investigate mechanisms and drug intervention for pressure overload-induced HF.

A simple enzyme-catalyzed reaction induced "switch" type fluorescence biosensor based on carbon nitride nanosheets for the assay of alkaline phosphatase activity.

An enzyme-catalyzed fluorescence "switch" type sensor was constructed for the determination of alkaline phosphatase (ALP) activity by combining the fluorescence quenching effect of Ag+ on ultrathin g-C3N4 nanosheets (CNNSs) with the simple redox reaction of AA and Ag+. Briefly, Ag+ exhibits a significant quenching effect on the fluorescence of CNNSs. Thus the fluorescence signal of the CNNS-Ag+ system is extremely weak even in the presence of l-ascorbic acid-2-phosphate (AAP) ("off" state). When ALP coexists in the system, the enzyme can specifically catalyze the hydrolysis of AAP to form ascorbic acid (AA), which reduces Ag+ to Ag0. In this case, the fluorescence signal of the system is recovered ("on" state). Based on this principle, a signal-enhanced CNNS fluorescence sensor was developed to determine the activity of alkaline phosphatase. The experimental results show that the detection range of alkaline phosphatase is 0.5-20 U L-1, and the detection limit is 0.05 U L-1 (S/N = 3). Meanwhile, this method was used to assay ALP in serum samples.

Masking quercetin: A simple strategy for selective detection of rutin by combination of bovine serum albumin and fluorescent silicon nanoparticles.

As a typical kind of bioactive flavonoid glycoside, rutin and its aglycone quercetin possess similar chemical structures and properties. It still remains a challenge to achieve reliably and accurately detection of rutin in the presence of quercetin. In this work, a simple fluorescent method combining water-dispersed silicon nanoparticles (SiNPs) with bovine serum albumin (BSA) were constructed for the selective detection of rutin in the presence of quercetin and other common compounds in traditional Chinese herbs. SiNPs with high fluorescent quantum yield and good thermostability were prepared by one-pot hydrothermal method using ferulic acid as the reduction reagent for the first time. The fluorescence of SiNPs could be obviously quenched both by rutin and quercetin in phosphate buffer solution. Interestingly, when the solution contained certain concentration of BSA, the fluorescence of the SiNPs can only be remarkably quenched by rutin. The innovative use of BSA to block the interference of quercetin make it possible to selectively detect of rutin by fluorescence spectrometry under the coexistence of quercetin. Under the optimum conditions, the fluorescence displayed a linear decrease response as the rutin concentration increased in the range of 0.33-33.30 µM with a detection limit of 0.04 µM (S/N = 3). The possible quenching mechanism of rutin to SiNPs has also explored and concluded to be mainly caused by inner filter effect. This work provides a novel methodology for the simple, low-cost and selective determination method for rutin.

Water-dispersed silicon quantum dots for on-off-on fluorometric determination of chromium(VI) and ascorbic acid.

Water-dispersed silicon quantum dots (SiQDs) with the quantum yield of 25% was prepared using aminopropyltrimethoxysilane as the silicon source and ascorbic acid (AA) as the reduction reagent. The SiQDs display blue fluorescence with excitation/emission peaks at 350 nm/440 nm. The synthesized SiQDs are shown to be a viable "on-off-on" fluorescent probe for the detection of Cr(VI) and AA. Cr(VI) ions exert an inner filter effect on the fluorescence of the SiQDs which results in a reduction of fluorescence (off-state). On addition of AA, Cr(VI) is chemically reduced to Cr(III) which weakens the inner filter effect and restores fluorescence (on-state). The method has low detection limits for both Cr(VI) and AA (0.16 µM and 0.57 µM, respectively). It was applied to the analysis of spiked lotus seeds and human serum samples. Graphical abstract A simple and facile "on-off-on" fluorometric method for Cr(VI) and ascorbic acid (AA) was developed using water-soluble silicon quantum dots (SiQDs) as the fluorescent probe. The approach was also used to assay Cr(VI) and AA in the lotus seeds and human serum, respectively.

Signal-on fluorescence assay for pyrophosphate ions based on DNA-stabilized silver nanoclusters.

Pyrophosphate anion (P2 O7 4- , PPi) is considered as a potential biomarker for arthritic diseases because high levels of PPi may result in calcium pyrophosphate dehydrate crystal deposition diseases. In this study, a simple fluorescence method for PPi was demonstrated by organic integration of the efficient fluorescence quenching ability of copper ions to DNA-scaffolded silver nanoclusters and the strong affinity of PPi towards copper ions. This simple fluorescence sensor showed a low detection limit (0.28 µM based on signal/noise = 3) towards the detection of PPi. Practical application of this method was also validated by detection of PPi in the synovial fluid.

Pien-Tze-Huang protects cerebral ischemic injury by inhibiting neuronal apoptosis in acute ischemic stroke rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Pien-Tze-Huang (PZH) is a famous formula of traditional Chinese medicine used to treating stroke. However, the protective effect of PZH and its mechanisms in acute ischemic stroke remain to be explored. AIM OF THE STUDY: To investigate the protective effect of PZH on neuronal apoptosis in acute cerebral ischemic injury rats and explore its underlying mechanisms. MATERIALS AND METHODS: The effects of PZH were studied in acute ischemic stroke rats induced by transient middle cerebral artery occlusion, and the mitochondria-mediated apoptotic proteins including cytochrome C (Cyt C), Bax, Bcl-xl, P53, caspase-3, and caspase-9 as well as AKT and glycogen synthase kinase-3 beta (GSK-3ß) were assessed. RESULTS: Four days of PZH treatment (180 mg/kg) could significantly reduce cerebral infarct volume, improve neurological deficit, attenuate inflammatory response, and inhibit neuronal apoptosis in acute ischemic stroke rats. Moreover, PZH treatment significantly decreased cytosolic Cyt C, Bax, P53, cleaved caspase-3, and cleaved caspase-9 levels, but elevated mitochondrial Cyt C and Bcl-xl levels. PZH treatment also increased phosphorylation of AKT and GSK-3ß. CONCLUSION: PZH potently protects the brain from cerebral ischemia/reperfusion injury in vivo, and inhibiting mitochondria-mediated neuronal apoptosis as well as attenuating inflammatory responses may be involved in this effect. This study provides experimental basis of PZH in treating acute cerebral ischemic stroke, which would provide some novel insights for its prevention and treatment of ischemic stroke.

Dispersive micro-solid-phase extraction combined with online preconcentration by capillary electrophoresis for the determination of glycopyrrolate stereoisomers in rat plasma.

A simple and sensitive analytical method for four isomers of glycopyrrolate in rat plasma was developed using cation-selective exhaustive injection-sweeping cyclodextrin-modified electrokinetic chromatography (CSEI-Sweeping-CDEKC) for online enrichment combined with dispersive micro-solid-phase extraction pretreatment. The CSEI-Sweeping-CDEKC was conducted on an uncoated fused silica capillary (40.2 cm × 75 µm) with an applied voltage of -20 kV. The electrophoretic analysis was carried out in 30 mM phosphate solution at pH 2.0 containing 20 mg/mL sulfated-ß-cyclodextrin and 5% acetonitrile. Under these optimized conditions, the detection limit for racemic glycopyrrolate was found to be 2.0 ng/mL and this method could increase 495-fold detection sensitivity compared with the traditional injection method. Additionally, the parameters that affected the extraction efficiency of dispersive micro-solid-phase extraction were also examined systematically. The glycopyrrolate isomers in rat plasma samples as low as 0.0625 µg/mL were able to be separated and detected by capillary electrophoresis with the aid of CSEI-sweeping. The findings of this study show that the dispersive micro-solid-phase extraction pretreatment coupled with CSEI-Sweeping-CDEKC is a rapid and convenient method for analyzing glycopyrrolate isomers in rat plasma.

Correlation between the presence of circulating tumor cells and the pathologic type and staging of non-small cell lung cancer during the early postoperative period.

This study investigated possible correlations between the presence of circulating tumor cells (CTCs) and the pathologic types and staging of non-small cell lung cancer (NSCLC) during the early postoperative period. Sixty-nine patients with NSCLC were enrolled in the study. Clinical staging was performed by postoperative pathological examination and imaging. Multiple mRNA in situ analyses targeting specifically expressed genes were carried out to identify the presence of CTCs. Correlations between age, sex, TNM stage and pathological types with the detection rate of CTCs were also established. The results showed the positivity rate of CTCs in patients >55 years was significantly higher than that of patients <55 years (94.74 vs. 70.97%, P<0.05). There was no significant difference in the positivity rate of CTCs between male and female patients (85.71 vs. 85.29%, P>0.05). The correlations between the detection rate of epithelial type or mixed type CTCs with tumor size, lymph node metastasis and distant metastasis TNM in patients with NSCLC were not significant (P>0.05). However, higher TNM stages correlated with higher detection rates of mesenchymal CTCs (P<0.05). There were also significant differences in the detection rates of CTCs amongst the three different pathologic types (adenocarcinoma, squamous cell and large cell carcinomas) (P<0.05). Based on our results, the detection of mesenchymal CTCs during the early postoperative period can help prognosticate the recurrence and metastasis of NSCLC, which is beneficial to the development of individualized treatment strategies.

Simultaneous Determination of 11 Compounds in Gualou Guizhi Granule and Pharmacokinetics Study by UPLC-MS/MS.

A rapid and sensitive ultrafast performance liquid chromatography-tandem mass spectrometry method (UPLC-MS/MS) was developed for the simultaneous determination of 11 compounds in Gualou Guizhi Granule (GLGZG), including liquiritin, isoliquiritin, liquirtin apioside, isoliquiritin apioside, liquiritigenin, isoliquiritigenin, glycyrrhizic acid, glycyrrhetinic acid, paeoniflorin, albiflorin, and paeoniflorin sulfonate in rat plasma. UPLC-MS/MS assay with negative ion mode was performed on a Waters CORTECS C18 (2.1 × 100 mm, 1.6 µm) with the mobile phase consisting of 0.1% aqueous formic acid (A) and acetonitrile (B) in gradient elution at a flow rate of 0.25 mL·min-1. The method was linear for all analytes within the detection range (r ≥ 0.9597). The inter- and intraday precision (RSD) were 2.21-6.41% and 1.67-6.18%; the inter- and intraday accuracy (recover) were 92.48-114.03% and 90.23-112.04%. And the recovery rate ranged from 81.30% to 108.22%. The matrix effect values obtained for analytes ranged from 88.91% to 113.32%. This validated method was successfully applied to a pharmacokinetics study in rats after oral administration of GLGZG.

Paeoniflorin exerts neuroprotective effects against glutamate­induced PC12 cellular cytotoxicity by inhibiting apoptosis.

Paeoniflorin (PF) is an active ingredient of Radix Paeoniae, which is known to exert neuroprotective effects. However, the mechanims behind the neuroprotective effects of PF are not yet fully understood. The apoptosis of neurons plays an important role in the cerebral ischemia-induced cascade response. This study aimed to investigate neuroprotective effects of PF against glutamate­induced PC12 cellular cytotoxicity and to determine whether these effects are mediated via the inhibition of apoptosis in vitro and the activity of mitochondrial apoptosis-associated proteins in PC12 cells. Exposure of the PC12 cells to glutamate induced cell morphological changes, significantly decreased cell viability and induced apoptosis, with similar results being observed from the Hoechst 33342 staining and Annexin V/PI staining experiments. Glutamate also increased the lactate dehydrogenase release by the PC12 cells. However, treatment with PF prevented these effects. Furthermore, PF inhibited Bax and Bad expression and increased Bcl-2 and Bcl-xL expression; it also decreased the levels of downstream protein (caspase-3 and caspase-9). Collectively, our results indicate that PF protects PC12 cells against glutamate-induced neurotoxicity possibly through the inhibition of the expression of mitochondrial apoptosis-associated proteins.

Application of Chiral Ionic Liquid-Modified Gold Nanoparticles in the Chiral Recognition of Amino Acid Enantiomers.

Two chiral ionic liquids (ILs), namely 1-ethyl-3-methylimidazole l-tartrate (EMIML-Tar) and 1-ethyl-3-methylimidazole l-lactate (EMIML-Lac), were used to modify gold nanoparticles (AuNPs) for chiral recognition of amino acid enantiomers. Transmission electron microscopy, infrared spectroscopy, ultraviolet-visible spectroscopy, and capillary electrophoresis were used for the characterization of chiral IL-modified AuNPs. Meanwhile, the performance of l-tartaric acid and l-lactic acid as modifiers was investigated to make a comparison. The chiral recognition mechanism is further discussed.

Extracts of Bauhinia championii (Benth.) Benth. attenuate the inflammatory response in a rat model of collagen-induced arthritis.

Rheumatoid arthritis is considered a serious public health problem, which is commonly treated with traditional Chinese or herbal medicine. The present study evaluated the effects of Bauhinia championii (Benth.) Benth. extraction (BCBE) on a type II collagen-induced arthritis (CIA) rat model. Wistar rats with CIA received either 125 or 500 mg/kg BCBE, after which, paw swelling was markedly suppressed compared with in the model group. In addition, BCBE significantly ameliorated pathological joint alterations, including synovial hyperplasia, and cartilage and bone destruction. The protein and mRNA expression levels of interleukin (IL)­6, IL­8, tumor necrosis factor­α and nuclear factor­κB in synovial tissue were determined by immunohistochemical staining, western blot analysis and reverse transcription­polymerase chain reaction. The results demonstrated that the expression levels of these factors were significantly downregulated in the BCBE­treated group compared with in the model group. These results indicated that BCBE may exert an inhibitory effect on the CIA rat model, and its therapeutic potential is associated with its anti-inflammatory action.

Extracts of Bauhinia championii (Benth.) Benth. inhibit NF-B-signaling in a rat model of collagen-induced arthritis and primary synovial cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Bauhinia championii (Benth.) Benth. is used in Chinese traditional medicine to treat arthritis, especially has been used a long time ago on rheumatoid arthritis (RA) in She ethnic minority group. AIM OF THE RESEARCH: To investigate the anti-RA effect of Bauhinia championii (Benth.) Benth ethyl acetate extract (BCBEE) and the molecular bases of it. MATERIALS AND METHODS: BCBEE was studied on a rat model of RA induced by Ⅱcollagen in vivo, as well as on primary synovial cells in vitro. RESULTS: After BCBEE treatment, in vivo, it was showed that paw and joint edema was inhibited, pathological joint changes was ameliorated and the levels of interleukin (IL)-1ß and tumor necrosis factor-(TNF-α) was decreased significantly. The protein and mRNA expressions of nuclear factor-B (NF-κB)(p65), IκB, p-IκB and IκB kinase beta (IκKß) were also down-regulated. Moreover, the in vitro study revealed that BCBEE treatment inhibited primary synovial cells proliferation, and promoted down-regulation of NF-κB(p65), IκB, p-IκB and IκKß. CONCLUSIONS: Taken together, the present study demonstrates that BCBEE produces a protection in a rat model of RA induced by Ⅱcollagen via inhibiting paw and joint edema, ameliorating pathological joint changes and regulating the levels of cytokines and its action mechanism maybe is via down-regulating NF-κB(p65), IκB, p-IκB and IκKß expression.

A bio-inspired sensor coupled with a bio-bar code and hybridization chain reaction for Hg(2+) assay.

In this article, a bio-inspired DNA sensor is developed, which is coupled with a bio-bar code and hybridization chain reaction. This bio-inspired sensor has a high sensitivity toward Hg(2+), and has been used to assay Hg(2+) in the extraction of Bauhinia championi with good satisfaction.

[Application of 1-ethyl-3-methyl imidazole L-lactate as the chiral ligand for the enantioseparation of amino acids].

Enantioseparation of tryptophan enantiomers (Trys), tyrosine enantiomers (Tyrs) and phenylamine enantiomers (Phes) by chiral ligand exchange capillary electrophoresis (CLE- CE) using 1-ethyl-3-methyl imidazole L-lactate ([EMIM] [L-Lac ]) as the chiral ligand was studied. The effects of background electrolytes and central ions, the molar ratio and concentration of the chiral ligand and central ion, running buffer solution pH on the enantioseparation of Trys, Phes and Tyrs were investigated. Good enantioseparation of three pairs of enantiomers were obtained with 40.0 mmol/L [EMIM] [L-Lac] and 20.0 mmol/L cupric chloride (pH 4.5). To validate the good behavior of [EMIM] [L-Lac], L-lactate (L-Lac) was further used as the chiral ligand for the enantioseparation of Trys. The contrast experiment showed that Trys could only be partially enantioseparated by L-Lac. Moreover, the enantioseparation was improved by adding 1-ethyl-3-methyl imidazole acetate (EMIM-Ace) which markedly prolonged the migra- tion time of Trys to more than 30 min. In contrast, the migration time of Trys was obtained in 15 min by [EMIM] [L-Lac]. Finally, the enantioseparation mechanism was further discussed. The experimental results showed that in the EMIM-Ace assisting L-Lac system, EMIM-Ace was only used to suppress the electroosmotic flow and was not involved in the chiral ligand exchange reaction, while in the [EMIM] [L-Lac] system, the chiral ligand taking part in the re- action was mainly unionized [EMIM] [L-Lac] rather than L-Lac formed by ionized L-Lac ions.

Label-free aptamer-based partial filling technique for enantioseparation and determination of DL-tryptophan with micellar electrokinetic chromatography.

In this study, a simple and reproducible method for enantioseparation and determination of dl-tryptophan (DL-Trp) was developed by using a partial filling technique in combination with MEKC. The corresponding L-Trp specific DNA aptamer was used as a chiral selector. Sodium cholate was used to form the chiral micelles and to enhance the enantioseparation of the enantiomers. Effects of aptamer concentration, filling time, buffer composition, and separation voltage on the enantioseparation were evaluated. The Mg(2+) and Na(+) concentration in separation buffer was found to effectively affect the separation efficiency and reproducibility. Under the optimal conditions, D- and L-Trp were completely enantioseparated in less than 9 min. This aptamer-based partial-filling approach has the potential to be extended to the separation of other enantiomers after the replacement of corresponding specific aptamers.

An ultrasensitive electrochemical impedance sensor for a special BRCA1 breast cancer gene sequence based on lambda exonuclease assisted target recycling amplification.

A label-free, target recycling electrochemical impedance spectroscopy (EIS) DNA sensor has been developed for detection of a model related to the BRCA1 breast cancer gene with a detection limit of 0.05 nM.