Targeting cancer cytoskeleton: the mechanical properties of natural anticancerdrugs

Anticancer drugs are one of the most direct means of cancer therapy.However,the various cancer progressions hamper the development and discovery of anticancer drugs.In fact,the mechanical properties of the tumor cytoskeleton are extremely vital for any phase of cancer,especially in tumor invasion and metastasis.However,in the current category of anticancer drugs,the cytoskeleton-targeting drugs are limited and their role in tumor progression is unclear.Here,we present the mechanical characteristics of tumor stiffness that are tightly regulated by the cancer cytoskeleton,including actin filaments and microtubules during tumor initiation,growth and metastasis,and review the natural drugs that target the cancer cytoskeleton.We define cytoskeleton dynamics as target mechanisms for anticancer drugs and summarize the plant,microbial and marine sources of natural products.Furthermore,this paper also provides a material pathway to study active tumor mechanics,and introduces the unique advantages and future application potential of tumor cytoskeleton-targeting drugs in clinical use.The material approaches to active cancer mechanics are supplied in this review.We aim to promote the development of anticancer drugs that target tumor mechanics by using those material approaches and finding their pharmacological application.

Comment on “A novel antibiotic class targeting the lipopolysaccharide transporter”

Crisis of antibiotic resistant has become a global threat to public health in recent decades[1–4].Carbapenem antibiotics are a class of atypicalβ-lactam antibiotics with the broadest antibacterial spectrum and the strongest antibacterial activity.However,carbapenem-resistant Acinetobacter baumannii(CRAB)is stilly emerging,which is a gram-negative bacterium with lipopolysaccharide(LPS),which makes it resistant to several antibiotics and thus difficult to eliminate[5,6].As the number one pathogen of nosocomial infection,CRAB can cause severe pneumonia and bloodstream infection,etc.The mortality rate of patients infected with invasive CRAB can be as high as 40%to 60%,and it has been listed as the key pathogen of Class 1 by the World Health Organization.Due to the lack of feasible antibiotic strategies,phage combination therapy has been used in some patients,while the efficacy is limited[7].Therefore,new discovery and development of antibiotic that targeting CRAB is urgently demand.Fortunately,Kenneth A.

Electroacupuncture improves microcirculation and neuronal morphology in the spinal cord of a rat model of intervertebral disc extrusion

Most studies on spinal cord neuronal injury have focused on spinal cord tissue histology and the expression of nerve cell damage and repair-related genes. The importance of the microcirculation is often ignored in spinal cord injury and repair research. Therefore, in this study, we established a rat model of intervertebral disc extrusion by inserting a silica gel pad into the left ventral sur-face of T13. Electroacupuncture was used to stimulate the bilateralZusanlipoint （ST36） and Neiting point （ST44） for 14 days. Compared with control animals, blood lfow in the ifrst lumbar vertebra （L1） was noticeably increased in rats given electroacupuncture. Microvessel density in the T13 segment of the spinal cord was increased significantly as well. The number of normal neurons was higher in the ventral horn of the spinal cord. In addition, vacuolation in the white matter was lessened. No obvious glial cell proliferation was visible. Furthermore, hindlimb motor function was improved signiifcantly. Collectively, our results suggest that electroacupuncture can improve neuronal morphology and microcirculation, and promote the recovery of neurological functions in a rat model of intervertebral disc extrusion.

Luteolin prevents f MLP-induced neutrophils adhesion via suppression of LFA-1 and phosphodiesterase 4 activity

Luteolin is an active ingredient found early from Fofium perillae and Flos Ionicerae, and has a specific inhibition on phosphodiesterase 4 （PDE4） activity in vitro. Researches show luteolin has pharmacological effects of anti-inflammation, anti-anaphylaxis, antitumor, antioxidant, protection of nervous system and so on, and has mainly been used for the treatment of respiratory inflammatory diseases, cancer and cardiovascular disease in clinic. PDE4, specific to hydrolyze cyclic AMP （cAMP）, is considered to be a new anti-inflammatory target due to the decisive role on cAMP signal in inflammatory cells such as neutrophils. In order to explore the anti-inflammatory mechanism, we further studied the effects of luteolin on the activity and expression of PDE4, the expression of lymphocyte function-associated antigen-1 （LFA-1） and macrophage-1 （MAC-l） in neutrophils, and the adhesion of neutrophils and endothelial cells. The results showed that luteolin had a dose-dependent inhibition on both bare PDE4 activity and PDE4 in cultured neutrophils, and had an obviously promotive effect on gene expressions of PDE4A, 4B and 4D in later period. Luteolin had a significant inhibitory effect on neutrophils adhesion and LFA-1 expression in early stage, and had no obvious effect on MAC-1 expression. Therefore, luteolin can inhibit LFA-1 expression of neutrophils, then inhibit the adhesion of neutrophils and endothelial cells, and the mechanism is at least related with the inhibition of PDE4 activity.

Influence of gallic acid on porcine neutrophils phosphodiesterase 4,IL-6,TNF-α and rat arthritis model

Our previous studies showed that the anti-inflammatory effects of Paeonia lactiflora roots extract may be mediated, at least in part, through its gallic acid content, and this effect may be regulated in part by an inhibition on c AMP-phosphodiesterase（PDE）. To explore the anti-inflammatory effect and mechanism, the influence of gallic acid on neutrophils PDE4 activity and expression, TNF-α and IL-6 content and rat arthritis model were further studied. PDE4 activity and gene express were calculated respectively by substrate c AMP change examined with HPLC and real-time RT-PCR. The concentration of IL-6 and TNF-α in supernatant were assayed by ELISA method. Model of rat arthritis was caused by complete Freund＇s adjuvant. Results showed that gallic acid had a dose-dependent restraint on PDE4 activity of neutrophils in vitro, promoted significantly PDE4 A expression（P〈0.01）, and had no influence on the expressions of PDE4 B and 4D. However, PDE4 C expression was not detected. Gallic acid could promote IL-6 release（P〈0.05）, and inhibit TNF-α release of neutrophils（P〈0.05）. The experiment in vivo showed that gallic acid had obvious restraint on local inflammation of animal model（P〈0.05）. Therefore, the anti-inflammatory effect of gallic acid may be mediated in part through an inhibition on PDE4 activity and further an increase of IL-6 and a decrease of TNF-α of neutrophils, and this effect seemed to have no relationship with PDE4 expression.

Effects of Polysaccharides from Pulsatilla Decoction on the Microvascular Endothelial Glycocalyx

Pulsatilla decoction is a famous traditional Chinese herbal formula for clearing heat and treating dysentery of animals or human. To elucidate its mechanism, many active components have been studied, however, the roles of its polysaccharides still remain unclear. This study aimed to explore effects of polysaccharides from Pulsatilla decoction （PPD） on the microvascular endothelial glycocalyx （eGC）. The polysaccharides were extracted from PPD by water extraction and alcohol precipitation method. Mice were administered with PPD for 4 wk, and were then anesthetized with ether inhalation and were ifxed by cardiac perfusion with gradient concentration alcian blue solution. The jejunum was sampled and jejunal mucosa was prepared for ultrathin sections by routine method and was analyzed by transmission electron microscope. The results indicated that the eGC was observed as a strong electron-dense smooth linear margin or nonuniform conglomerates coating cell membranes, and PPD signiifcantly increased its thickness from （21.85±1.87） to （28.71±3.61） nm and improved its integrity. This study suggested that PPD may express their biological activities and protect against pathogenic factor damages by inlfuencing the eGC.

Differences Between the Concentrations in Rabbit Body Fluid of Berberine Hydrochloride, Chlorogenic Acid, Baicalin, and Their Minimal Inhibitory Concentrations

To explore the relationship between the heat-clearing and detoxicating functions and the bacteriostatic actions of berberine hydrochloride （Ber. H）, chlorogenic acid （Chlo. A）, and baicalin （Bai）, their concentrations in rabbit body fluid were compared with their minimal inhibitory concentrations （MICs）. Their concentrations in rabbit blood and tissue fluid were determined by reversed-phase high performance liquid chromatography, and their MICs to Escherichia coli were determined by tube dilution method. The results showed that the peak concentrations of Ber. H, Chlo. A, and Bai in rabbit blood were 3.2, 5.03, and 7.63 μg mL^-1, and in rabbit tissue fluid were 0.12, 0.11, and 0.12 μg mL^-1, respectively. Their MICs to E. coli were, respectively, 1.0×10^3, 3.75 × 10^3, and 6.75 ×10^3μg mL^-1, which were far higher than the concentrations in rabbit body fluids. This study indicates that Ber. H, Chlo. A, and Bai have weak bacteriostatic actions and do not reach their effective inhibitory concentrations in rabbit body fluids, and their heat-clearing and detoxicating functions are independent on the bacteriostatic actions.

Synthesis and Metabolism of 5-hydroxytryptamine in Peripheral Circulatory System of Suckling Piglets

[ Objective] To find the rule of 5-hydroxytryptamine （5-HT） synthesis and metabolism in peripheral blood of suckling piglets with aging. [ Method ] All of 15 British Large White pigs at 0, 5 and 15 d of age were selected as experimental animals, and their serumal concentrations of L- tryptophan （L-Trp）, 5-HT and 5-hydroxyindoleacetic acid （5-HIAA） in peripheral circulatory system were determined synchronously by reversed- phase high-performance liquid chromatography method with UV. [ Result] The concentration of L-Trp was （16. 92 ± 1.74） iJmol/L at birth, （59.94 ±10.88） iJmol/L at 5 d of age and （70.42± 6.48） μmol/L at 15 d of age. The concentration of 5-HT was （ 12.85 ± 1.79） iJmol/L at birth, （4.81 ±2.05） ijmol/L at 5 d of age and （5.49±1.09） μmol/L at 15 d of age. While 5-HIAA, the metabolic product of 5-HT, was not detected at all the samples selected. [ Conclusion] The L-Trp concentration of suckling piglets increase significantly with aging （ P 〈0. 05）. While the concentra- tion of 5-HT and the conversion ratio of L-Trp to 5-HT are both highest at birth （P〈0.05） and keep stable at 5 and 15 d of age.

Effects of Ginsenoside Rb1 Babu Agent on the Amplitude of Microvascular Vasomotion

[Objectives] To study the effects of ginsenoside Rbl Babu agent on the amplitude of microvascular vasomotion. [Methods]The in vitro transdermal permeation of ginsenoside Rbl Babu agent was performed by using intelligent transdermal apparatus,and the percutaneous fluid was collected and analyzed by using high performance liquid chromatography( HPLC). The Babu agent was stuck on the skin of acupoint area,determined by Laser Doppler Flowmeter,and amplitude of microvascular vasomotion of acupoint area was recorded. [Results] With the extension of transdermal time,the cumulative permeation rate of ginsenoside Rbl increased. The amplitude of microvascular vasomotion could be significantly increased with the application of ginsenoside Rbl Babu agent( P < 0. 01). [Conclusions]The drug delivery system of ginsenoside Rbl Babu agent can release the drug into the acupoint,increase the amplitude of microvascular vasomotion,and achieve the effect of acupuncture. Therefore,ginsenoside Rbl Babu agent can replace the acupuncture clinically to treat diseases.

Detection of Enterocin Gene from Enterococcus

[ Objective] This study aimed to clone and identify enterocin gene from Enterococcus. [ Method] The genomic DNA of 12 enterocecci was extracted, and separately amplified with specific primers. The amplified fragments were ligated into PGEM-T Easy vector, which was then transformed into DH5α competent cells. The positive clones were sequenced. [Result] Enterocin A gene was 274 bp long. It was obtained from six enterococci, and the amino acids encoded by the enterocin genes cloned from five object enterocecci were the same as that of type IIa reference strains except only one amino acid. The homology among them reached 99.76 - 100%, suggesting that the bacteriocin isolated from the enterococcis belonged to type II. Structure prediction by DNAstar indicated that 22nd - 30th amino acids of enterocin A formed ot region, which had a hydrophilic region at its N-terminal and a hydrophobic region at its C-terminal, a transmembrane helix structure. [ Conclusion] This study will provide basis for the heterologous expression and applications of enterocins. Key words Enterocin gene; Enterocecci; PCR; Sequence analysis

Lactic Acid Reduces LPS-Induced TNF-a and IL-6 mRNA Levels Through Decreasing I?Ba Phosphorylation

This study explored the effects over time of lactic acid （LA） on IκBα phosphorylation and nuclear factor-kappa B （NF-κB） p65 protein expression, and on tumor necrosis factor a （TNF-α） and interleukin-6 （IL-6） mRNA levels in rat intestinal mucosa microvascular endothelial cells （RIMMVECs） stimulated by lipopolysaccharide （LPS）. I？Ba, phosphorylated IκBa （p-IκBa） and p65 protein levels were monitored by Western blot analysis, and TNF-a and IL-6 mRNA levels were analyzed using real-time PCR. LA treatment reduced TNF-a and IL-6 mRNA levels in LPS-stimulated RIMMVECs, with the greatest effect being after 3 h. The highest inhibitory effect of LA on IκBa phosphorylation to prevent activation of NF-κB was after 6 h. These results suggest that LA reduces TNF-a and IL-6 mRNA levels through decreasing IκBa phosphorylation and blocking the dissociation of IKK complex, which prevents activation of NF-κB.

Baicalin Induces IFN-α/β and IFN-γ Expressions in Cultured Mouse Pulmonary Microvascular Endothelial Cells

We studied the effect of baicalin,an extract from Radix Scutellariae（a traditional Chinese medicine） in inducing mouse pulmonary microvascular endothelial cells（MPMVECs） to produce interferons（IFNs）.MPMVECs were cultured in vitro in the presence of different concentrations of baicalin（10,20,and 30 μg mL-1）,and the cells and the culture media were harvested at various time intervals.The proteins and mRNA levels（relative to β-actin） of IFN-α/β and IFN-γ were analyzed by RT-PCR and enzyme-linked immunosorbent assay（ELISA）.It was observed that baicalin substantially up-regulated the expression of IFN-α/β and IFN-γ.In all baicalin-treated groups,the relative levels of IFN-α/β and IFN-γ mRNAs peaked after 12 h of culturing,and IFN-α/β and IFN-γ proteins peaked after 24 h of culturing.These results suggest that baicalin can effectively induce the expression of IFNs in pulmonary microvascular endothelial cells,and thus potentially act as an antiviral compound.This study may provide background information for developing new antiviral drugs based on baicalin.

Electroacupuncture improves neuronal function by stimulation of ascending peripheral nerve conduction in rats with spinal cord injury

OBJECTIVE: To establish a method of transient sciatic nerve blockade and to examine the involvement of the ascending peripheral nerve pathway in the therapeutic effect of electroacupuncture at Zusanli(ST 36) in rats with spinal cord injury(SCI).METHODS: We examined the transient effect of daily lidocaine administration into the posteromedial part of the greater trochanter on sciatic nerve function using electrophysiological examination and histopathology of the sciatic nerve. Rats were divided into three groups: an SCI group(SCI without treatment), an SCI with electroacupuncture treatment(SCI-EA) group, and an SCI with nerve block and electroacupuncture(SCI-NB-EA) group(nerve block was achieved by lidocaine administration to transiently block the ascending peripheral nerve pathway). Behavioral tests and electrophysiological examinations were performed to evaluate recovery of neurological function.RESULTS: Sciatic nerve conduction was normal immediately before daily lidocaine administration.Histopathological analysis also indicated normal sciatic nerve, confirming that lidocaine nerve blockade was suitable and reversible for transiently eliminating nerve transmission. Neurological function in the SCI-EA group was superior to that in the SCI group, while no differences were found between the SCI and SCI-NB-EA groups.CONCLUSION: Electroacupuncture treatment can promote recovery of neurological function. Facilitation of nerve conduction may play an important role in this recovery.

Capillary blood flow in patients with dysmenorrhea treated with acupuncture

OBJECTIVE: To observe capillary blood flow at acupoints during acupuncture treatment of primary dysmenorrhea and gain new insights into its analgesic mechanism. METHODS: Patients with primary dysmenorrhea were enrolled and randomly assigned to a treatment or control group. Subjects' symptoms were differentiated into variousTraditional Chinese Medicine(TCM) syndromes and treated for 10 sessions with puncturing acupuncture or self-pressing right-hand Hegu(LI 4), adding other acupoints based on syndrome. Laser speckle was used to compare the change in the vasomotor amplitude and perfusion of the capillaries in Hegu(LI 4) before and during the treatment. Each subject was required to finish the period pain symptoms observation form, verbal rating scales, numerical rating scale, pain rating index, face rating scale, Zung self-rating depression scale, Zung self-rating anxiety scale, and numerical rating scale before and after treatments. RESULTS: After 10 sessions, the symptom scores, pain index(PI), and visual analog scale(VAS) decreased significantly in treatment group. The volume of blood flow in Hegu(LI 4) declined slightly. No significant evidence supported that needling caused capillary contraction, but the capillary vasomotor amplitude at Hegu(LI 4) increased remarkably. CONCLUSION: Acupuncture can increase the capillary blood flow, thus promoting the flow of Qi and blood in terms of TCM theory, which facilitates pain relief.