Cancer nutritional-immunotherapy with NIR-II laser-controlled ATP release based on material repurposing strategy.

Enlightened by the great success of the drug repurposing strategy in the pharmaceutical industry, in the current study, material repurposing is proposed where the performance of carbonyl iron powder (CIP), a nutritional intervention agent of iron supplement approved by the US FDA for iron deficiency anemia in clinic, was explored in anti-cancer treatment. Besides the abnormal iron metabolic characteristics of tumors, serving as potential targets for CIP-based cancer therapy under the repurposing paradigm, the efficacy of CIP as a catalyst in the Fenton reaction, activator for dihydroartemisinin (DHA), thus increasing the chemo-sensitivity of tumors, as well as a potent agent for NIR-II photothermal therapy (PTT) was fully evaluated in an injectable alginate hydrogel form. The CIP-ALG gel caused a rapid temperature rise in the tumor site under NIR-II laser irradiation, leading to complete ablation in the primary tumor. Further, this photothermal-ablation led to the significant release of ATP, and in the bilateral tumor model, both primary tumor ablation and inhibition of secondary tumor were observed simultaneously under the synergistic tumor treatment of nutritional-photothermal therapy (NT/PTT). Thus, material repurposing was confirmed by our pioneering trial and CIP-ALG-meditated NT/PTT/immunotherapy provides a new choice for safe and efficient tumor therapy.

Scalp Acupuncture Enhances the Functional Connectivity of Visual and Cognitive-Motor Function Network of Patients with Acute Ischemic Stroke.

DESIGN: A parallel-group randomized controlled trial. Participants. 30 hemiplegic patients with middle cerebral artery acute infarction of the dominant hemisphere. Interventions. 30 patients were divided into 2 groups randomly. 15 patients in the treatment group (TG) were treated with ISSA, needling at the parietal midline (MS5) and left anterior/posterior parietal-temporal oblique lines (MS6 and MS7), combined with western routine treatment. While another 15 patients in the control group (CG) received routine treatment only. Main Outcome Measures. (1) Functional connectivity (FC): patients received brain scan using 3.0 T MRI after the treatment for 1 week. Based on the Matlab2012a platform, SPM12 software and DPABI software were used to process the scanning data and finally the functional connectivity of the brain was obtained. (2) National Institute of Health Stroke Scale (NIHSS) score. RESULTS: The difference in the NIHSS score between the two groups of patients before and after treatment was statistically significant (tNIHSS = 2.225; PNIHSS = 0.038), indicating that TG had a better effect. Centered to the seed region of the left supplementary motor area (SMA) (-5.32, 4.85, 61.38), FC increased at the left middle cerebellar peduncle, left cerebellum posterior lobe (uvula and declive), vermis, fusiform gyrus, lingual gyrus, inferior occipital gyrus, calcarine, cuneus, precuneus, BA7, BA18 and BA19, etc. Centered to the seed region of the left parahippocampal gyrus (PG) (-21.17, -15.95, -20.70), FC increased at the left precuneus, inside-paracingulate, inferior parietal gyrus, paracentral lobule, BA5, BA6, BA7, and BA40, right median cingulate, precuneus, BA19, BA23, and BA31, etc. CONCLUSIONS: It is indicated that ISSA can regulate the brain functional connection in patients with middle cerebral artery acute infarction in the dominant hemisphere and specifically strengthen the connections between visual, cognitive, motor control, and planning-related brain regions, which may be related to the recovery of movement in the mechanism. This trial is registered with ChiCTR-IOR-15007672.

Non-Magnetic Injectable Implant for Magnetic Field-Driven Thermochemotherapy and Dual Stimuli-Responsive Drug Delivery: Transformable Liquid Metal Hybrid Platform for Cancer Theranostics.

Transformable liquid metal (LM)-based materials have attracted considerable research interest in biomedicine. However, the potential biomedical applications of LMs have not yet been fully explored. Herein, for the first trial, the inductive heating property of gallium-indium eutectic alloy (EGaIn) under alterative magnetic field is systematically investigated. By virtue of its inherent metallic nature, LM possesses excellent magnetic heating property as compared to the conventional magnetite nanoparticles, therefore enabling its unique application as non-magnetic agents in magnetic hyperthermia. Moreover, the extremely high surface tension of LM could be dramatically lowered by a rather facile PEGylation approach, making LM an ideal carrier for other theranostic cargos. By incorporating doxorubicin (DOX)-loaded mesoporous silica (DOX-MS) within PEGylated LM, a magnetic field-driven transformable LM hybrid platform capable of pH/AFM dual stimuli-responsive drug release and magnetic thermochemotherapy are successfully fabricated. The potential application for breast cancer treatment is demonstrated. Furthermore, the large X-ray attenuation ability of LM endows the hybrid with the promising ability for CT imaging. This work explores a new biomedical use of LM and a promising cancer treatment protocol based on LM hybrid for magnetic hyperthermia combined with dual stimuli-responsive chemotherapy and CT imaging.

Identification of Ophiocordyceps sinensis and Its Artificially Cultured Ophiocordyceps Mycelia by Ultra-Performance Liquid Chromatography/Orbitrap Fusion Mass Spectrometry and Chemometrics.

Since the cost of Ophiocordyceps sinensis, an important fungal drug used in Chinese medicine, has increased dramatically, and the counterfeits may have adverse health effects, a rapid and precise marker using the peptide mass spectrometry identification system could significantly enhance the regulatory capacity. In this study, we determined the marker peptides in the digested mixtures of fungal proteins in wild O. sinensis fruiting bodies and various commercially available mycelium fermented powders using ultra-performance liquid chromatography/Orbitrap Fusion mass spectrometry coupled with chemometrics. The results indicated the following marker peptides: TLLEAIDSIEPPK (m/z 713.39) was identified in the wild O. sinensis fruiting body, AVLSDAITLVR (m/z 579.34) was detected in the fermented O. sinensis mycelium powder, FAELLEK (m/z 849.47) was found in the fermented Ophiocordyceps mycelium powder, LESVVTSFTK (m/z 555.80) was discovered in the artificial Ophiocordyceps mycelium powder, and VPSSAVLR (m/z 414.75) was observed in O. mortierella mycelium powder. In order to verify the specificity and applicability of the method, the five marker peptides were synthesized and tested on all samples. All in all, to the best of our knowledge, this is the first time that mass spectrometry has been employed to detect the marker peptides of O.sinensis and its related products.

Long-term acupuncture treatment has a multi-targeting regulation on multiple brain regions in rats with Alzheimer's disease: a positron emission tomography study.

The acute effect of acupuncture on Alzheimer's disease, i.e., on brain activation during treatment, has been reported. However, the effect of long-term acupuncture on brain activation in Alzheimer's disease is unclear. Therefore, in this study, we performed long-term needling at Zusanli (ST36) or a sham point (1.5 mm lateral to ST36) in a rat Alzheimer's disease model, for 30 minutes, once per day, for 30 days. The rats underwent 18F-fluorodeoxyglucose positron emission tomography scanning. Positron emission tomography images were processed with SPM2. The brain areas activated after needling at ST36 included the left hippocampus, the left orbital cortex, the left infralimbic cortex, the left olfactory cortex, the left cerebellum and the left pons. In the sham-point group, the activated regions were similar to those in the ST36 group. However, the ST36 group showed greater activation in the cerebellum and pons than the sham-point group. These findings suggest that long-term acupuncture treatment has targeted regulatory effects on multiple brain regions in rats with Alzheimer's disease.

Acupuncture at KI3 in healthy volunteers induces specific cortical functional activity: an fMRI study.

BACKGROUND: Using functional magnetic resonance imaging (fMRI), we determined brain regions that were activated/deactivated more by acupuncture at Taixi (KI3) than by non-acupoint or sham acupuncture. METHODS: A total of 30 healthy volunteers were randomly divided into a KI3 group (15 subjects) and non-acupoint group (15 subjects). Subjects in KI3 group received a sham acupuncture and then a real acupuncture, fMRI was performed before and after sham acupuncture as well as after ture acupuncture. Subjects in non-acupoint group received a ture acupuncture and the fMRI was performed before and after ture acupuncture. The fMRI data obtained were successively analyzed using DPARSF2.3 and REST1.8 software, yielding regional homogeneity (ReHo) and amplitude of low frequency fluctuations (ALFF) values. RESULTS: Compared with sham acupuncture, ALFF values were higher in Brodmann area (BA) 10 and lower in BA7 and BA18. ReHo values after real acupuncture at KI3 were higher in the right sub-lobar region and BA10 and were lower in BA31. Compared with the changes before and after real acupuncture at non-acupoint, the changes at KI3 showed higher ALFF valued in the left cerebellum posterior lobe, BA10, BA39, BA31 and decreased ALFF was observed in the BA18, BA19 and BA40; and higher ReHo values were shown in left cerebellum posterior lobe pyramis, left cerebellum anterior lobe. BA37, BA10, BA39, BA31 and lower ReHo values were shown in BA18 and BA31. CONCLUSION: Acupuncture at KI3 has a specific effect on certain brain regions associated with perception, body movement, spirit, and association. Additionally, visual and auditory cortices were affected, which may be related to the clinical applications of KI3 acupuncture in auditory and cognitive disorders, hypomnesis, loss of concentration, and the loss of ability to work and learn. TRIAL REGISTRATION: The research ethics committee was achieved at 01/08/2012, the NO. was ChiECRCT-2012011. Website for Clinical Trial Registration: http://www.chictr.org.cn/showproj.aspx?proj=7123 . This study was registered at www.chictr.org, the Clinical Trial Registration Number was ChiCTR-TRC-12002427, and the registration number was achieved at 18/08/2012. The name of IRB that provided approval for the study and clearly state is Chinese Clinical Trail Registry.

Acupuncture for ischemic stroke: cerebellar activation may be a central mechanism following Deqi.

The needling sensation of Deqi during acupuncture is a key factor of influencing acupuncture outcome. Recent studies have mainly focused on the brain function effects of Deqi in a physiological state. Functional magnetic resonance imaging (fMRI) on the effects of acupuncture at Waiguan (SJ5) in pathological and physiological states is controversial. In this study, 12 patients with ischemic stroke received acupuncture at Waiguan (SJ5) and simultaneously underwent fMRI scanning of the brain, with imaging data of the activated areas obtained. Based on the patient's sensation, imaging data were allocated to either the Deqi group or non-Deqi group. In the Deqi group, the activated/deactivated areas were the left superior temporal gyrus (BA39)/right anterior lobe of the cerebellum and left thalamus. In the non-Deqi group, the activated areas included the medial frontal gyrus of the right frontal lobe (BA11), right limbic lobe (BA30, 35), and left frontal lobe (BA47), while the only deactivated area was the right parietal lobe (BA40). Compared with the non-Deqi group, the Deqi group exhibited marked activation of the right anterior lobe of the cerebellum and right limbic lobe (BA30). These findings confirm that the clinical effect of Deqi during acupuncture is based on brain functional changes. Cerebellar activation may be one of the central mechanisms of acupuncture in the treatment of ischemic stroke.

Extraction, purification and antioxidant activities of the polysaccharides from maca (Lepidium meyenii).

Water-soluble polysaccharides were separated from maca (Lepidium meyenii) aqueous extract (MAE). The crude polysaccharides were deproteinized by Sevag method. During the preparation process of maca polysaccharides, amylase and glucoamylase effectively removed starch in maca polysaccharides. Four Lepidium meyenii polysaccharides (LMPs) were obtained by changing the concentration of ethanol in the process of polysaccharide precipitation. All of the LMPs were composed of rhamnose, arabinose, glucose and galactose. Antioxidant activity tests revealed that LMP-60 showed good capability of scavenging hydroxyl free radical and superoxide radical at 2.0mg/mL, the scavenging rate was 52.9% and 85.8%, respectively. Therefore, the results showed that maca polysaccharides had a high antioxidant activity and could be explored as the source of bioactive compounds.

Key regions of the cerebral network are altered after electroacupuncture at the Baihui (GV20) and Yintang acupuncture points in healthy volunteers: an analysis based on resting fcMRI.

OBJECTIVE: To identify the key cerebral functional region affected by acupuncture point needling by examining cerebral networks using functional connectivity MRI (fcMRI) and analysing changes in the key regions of these brain networks at different time points after needle removal. METHODS: Twelve healthy volunteers received 30 min of electroacupuncture (EA) at the Baihui (GV20) and Yintang acupuncture points and then underwent two fMRI scans, one each at 5 and 15 min after needle removal. Related brain networks were analysed centred at different 'seeds', centres which functionally connect the other cerebral regions in an organised network, such as the anterior frontal lobe, anterior cingulate gyrus, parahippocampal gyrus, amygdala, hypothalamus, head of the caudate nucleus and anterior lobe of the cerebellum. Networks were analysed based on the resting cerebral functional connection, and the differences in the activities of the brain networks between the two time points were compared. RESULTS: At 5 min after needle removal, 12 brain functional regions were involved in organising the network centred at the caudate nucleus 'seed.' This number was greater than the number of related brain networks centred at the other 'seeds'. At 15 min after needle removal, 15 and 14 brain functional regions were involved in organised networks centred at the parahippocampal and hypothalamus 'seeds', respectively; these numbers were greater than the numbers of other related brain networks centred at the other 'seeds'. CONCLUSIONS: A brain network composed of a large number of cerebral functional regions was found after EA at GV20 and Yintang in healthy volunteers. The key brain 'seed' supporting the largest brain network changed between 5 and 15 min after needle removal.

[Isolation and mass spectrometric analysis of antioxidant peptides from enzymatic hydrolysates of roasted pills of Asini Corii Colla].

A fraction named GFC-1 with high antioxidant activities in vitro was isolated from the enzymatic hydrolysates of roasted pills of Asini Corii Colla, and the peptides in this fraction were identified. The enzymatic hydrolysates were isolated and purified with anion exchange chromatography and Sephadex G-25 filtration chromatography successively. GFC-1, a fraction isolated from the hydrolysates, exhibited the highest DPPH and ABTS scavenging capacity (DPPH 47. 95% at 2.0 g x L(-1) and ABTS 97.20% at 0.40 g x L(-1). Nine peptides from GFC-1 were identified by LC-ESI-MS/MS coupled with TurboSEQUEST search software and Swiss-Prot data base, and a high repetition core sequence GPAGPP*GPP* was also found.

[Peptides analysis in digested edible bird's nest by HPLC-MS].

Edible bird's nest contains lots of glycoproteins. The glycosylation inhomogeneity for glycoprotein often results in wide range of molecular weight and the difficulty for protein separation and charaterization. In this paper, proteins in the edible bird's nest were extracted using multiple extractions, and then digested by PNgase F and trypsin. The digest mixture was separated with HPLC, and peptides were identified based on MS/MS data searching. The results indicated that the extracted proteins were amount to 79.7% of total protein in the edible bird's nest. More than 20 species of peptides in the digested mixture were identified. The sequences of these peptides showed similarity with some proteins from Swiss-prot. The research indicated that deglycosylation, tryptic digestion coupled with HPLC-MS/MS is a proper strategy for characterization of proteins in the edible bird's nest.

Brain areas involved in acupuncture needling sensation of de qi: a single-photon emission computed tomography (SPECT) study.

BACKGROUND: De qi is a sensory response elicited by acupuncture stimulation. According to traditional Chinese medicine (TCM), de qi is essential for clinical efficacy. However, the understanding of the neurobiological basis of de qi is still limited. OBJECTIVE: To investigate the relationship between brain activation and de qi by taking a single-photon emission computed tomography (SPECT) scan while applying acupuncture at TE5. METHODS: A total of 24 volunteers were randomly divided into 4 groups, and received verum or sham acupuncture at true acupuncture point TE5 or a nearby sham point according to grouping. All subjects then received a (99m)Tc-ethylcysteinate dimer (ECD) SPECT scan. RESULTS: All six subjects in the verum acupuncture at true acupuncture point group experienced de qi sensation; in contrast, all six subjects in the sham acupuncture at the sham point group responded with nothing other than non-sensation. Compared to the scan results from subjects who experienced non-sensation, SPECT scans from subjects with de qi sensation demonstrated significant activated points mainly located in brodmann areas 6, 8, 19, 21, 28, 33, 35, 37, 47, the parahippocampal gyrus, lentiform nucleus, claustrum and red nucleus; deactivated points were seen in brodmann areas 9 and 25. CONCLUSIONS: Verum acupuncture at true acupuncture points is more likely to elicit de qi sensation. De qi sensations mainly resulted in brain area activations, but not deactivations. These brain areas are related to the curative effect of Te5. The acupuncture needle sensations of de qi and sharp pain are associated with different patterns of activations and deactivations in the brain.

[Characteristics of PET cerebral functional imaging during "Deqi" of acupuncture in healthy volunteers].

OBJECTIVE: To observe the characteristics of needling sensation of "Deqi" (feelings of soreness, numbness, distending and heaviness, SNDH) by using positron emission tomography (PET) based on changes of glucose metabolism in different functional brain areas. METHODS: Eighteen normal volunteers [9 men and 9 women, mean age (23.23-1- 3. 32) years] were randomly divided into control, Waiguan (SJ 5) and non-acupoint groups (n=6 in each group). SJ 5 and non-acupoint (the midpoint between SJ 5 and the running course of the Small Intestine Meridian on the right forearm) were punctured by using a sterilized filiform needle. PET scan of the brain began 40 min after intravenous 18 F-fluorodeoxyglucose (FDG) injection (0. 11 mCi/kg body weight, left opisthenar vein). Needling sensations including "Deqi"(n= 5), tingling (n 5) and no-apparently-specific-feeling (NASF) were acquired by acupuncture stimulation and grouped. The needling sensations were evaluated by using Visual Analog Scale(VAS). The acquired image data of different needling-sensation groups were analyzed using SPM 2. 0 software in the Matlab Platform. RESULTS: After receiving acupuncture stimulation of SJ 5, 5 volunteers in the Waiguan (SJ 5) group experienced fee- lings of SNDH, with the mean VAS score being 4.23 +/- 1. 50, and 5 volunteers of the non-acupoint group had a tingling feeling, with the mean VAS score being 5.73 2.40. The VAS score of the tingling group was significantly higher than that of SNDH group (P<0. 05). Compared with the NASF control group, the activated cerebral areas were Brodmann area (BA) 7, 13, 20, 22, 39, 42 and BA 45 in the SNDH group, mainly involving the left temporal lobe, superior temporal gyrus, etc. and being obviously different to those of the control group (P<0. 001,k>10 voxels). The activated cerebral areas in the tingling group were BA 18, 19, 22, 24, 25, 32, 36, 40 and BA 45, predominantly involving the left limbic lobe, hippocampal gyrus, etc. and being apparently different to those of the control group (P<0. 001,k> 10 voxels). CONCLUSION: Acupuncture of Waiguan (SJ 5) mainly produces feelings of soreness, numbness, distending and heaviness, and the activated cerebral areas mainly involve the left temporal lobe, superior temporal gyrus, etc. ; while acupuncture of its neighboring non-acupoint chiefly induces a feeling of tingling in the healthy subjects, and the activated regions predominately involve the left limbic lobe, hippocampal gyrus, etc.

[Investigation on degradation process of collagens during manufacturing procedure of E'jiao].

OBJECTIVE: To investigate the degradation process of collagens and identify the key unit operation during manufacturing process of E'jiao. METHOD: Samples in different unit operations were withdrawn, and their amino acid compositions and the molecular weight ranges were determined. The peptide composition was analyzed by high-performance liquid chromatography/mass spectrometry. RESULT: The content of sample during atmospheric condensation unit increased by 16.8% compared to the thermal extraction unit. Gel filtration chromatographic analysis indicated that the degradation process of collagen primarily occurred during the atmospheric condensation unit. The peptides in samples mainly resulted from the degradation of collagens and cytoskeleton proteins such as tubulin, actinin, and so on. The relative abundance of degraded collagens increased with the decrease of no-collagen proteins. CONCLUSION: Collagen degradation mainly occurred during the atmospheric condensation unit, which was the key process affecting the composition of E-jiao.

A cerebral functional imaging study by positron emission tomography in healthy volunteers receiving true or sham acupuncture needling.

Our recent studies have demonstrated that needling in Baihui, Shuigou and Shenmen enhances glucose metabolism in the frontal lobes, thalamus, temporal lobe, and the lentiform nucleus in vascular dementia. This study examined the effect of true, sham and overt needling in Waiguan (TE5) on cerebral changes by positron emission tomography (PET) technique. Eighteen healthy volunteers were randomized to receive overt control, true or sham needling therapy. To manipulate true needling, a needle was inserted into 15+/-2mm into Waiguan and "deqi" was achieved by proper needle manipulation. For sham needling, needles with a blunt tip were pushed against the skin as the shaft moved into the handle, giving an illusion of insertion. For overt placebo, blunt needles were used and subjects did not receive any needling penetration. The tracer used was (18)fluoride-deoxygluocse. PET images obtained were processed and analyzed by the SPM2 software. Compared with overt needling, brain areas BA7, 13, 18, 19, 21, 22, 27, 38, 40, 42 and 45 in Waiguan true needling group were significantly activated and areas BA4, 6, 7, 19, 22 and 41 in sham needling group showed obvious activation. Compared to sham needling group, marked activation points were found in the areas of BA13 and 42 and left cerebellum in true needling group. Our study revealed a marked difference in brain metabolic changes between true and sham needling in Waiguan. Further studies are needed to explore the cerebral changes in patients with acupuncture and the pathological implications.

One-step purification of epigallocatechin gallate from crude green tea extracts by isocratic hydrogen bond adsorption chromatography on beta-cyclodextrin substituted agarose gel media.

An oligomerized beta-cyclodextrin ligand coupled to brominated allyl-group substituted Sepharose HP has been used for the one-step purification of polyphenolic epigallocatechin gallate (EGCG), an important antioxidant, by isocratic hydrogen bond adsorption chromatography. With a sample load of 1.33 mg crude green tea polyphenolic extract per ml column packing and with water/ethanol/acetonitrile (57/30/13, v/v) as the optimum mobile phase, an EGCG purity of about 98% with a recovery of approximate 73% could be achieved by proper peak cutting. After about 10 sample applications, the column performance started to deteriorate but could be regenerated to its original function by cleaning with 0.35 M NaOH.

Identification of major active constituents in the fingerprint of Salvia miltiorrhiza Bunge developed by high-speed counter-current chromatography.

High-speed counter-current chromatography was applied as a method to develop fingerprinting of Salvia miltiorrhiza Bunge, a popular traditional Chinese medicine, in our previous study. Important active constituents that were directly related to the therapy effect should be identified. Each effluent fraction and standard samples (cryptotanshinone, tanshinoneI and tanshinoneIIA) were analyzed by ultraviolet spectroscopy and liquid chromatography-mass spectroscopy. It was concluded from the UV-Vis spectrograms, retention times in LC analysis and mass spectrograms, that fractions 7, 8 and 11 were respectively cryptotanshinone (Mr 296), tanshinone I (Mr 276) and tanshinone IIA (Mr 294).