Vehicle-Free Nanotheranostic Self-Assembled from Clinically Approved Dyes for Cancer Fluorescence Imaging and Photothermal/Photodynamic Combinational Therapy.

Phototherapy, including photothermal therapy (PTT) and photodynamic therapy (PDT) has attracted growing attention as a noninvasive option for cancer treatment. At present, researchers have developed various "all-in-one" nanoplatforms for cancer imaging and PTT/PDT combinational therapy. However, the complex structure, tedious preparation procedures, overuse of extra carriers and severe side effects hinder their biomedical applications. In this work, we reported a nanoplatform (designated as ICG-MB) self-assembly from two different FDA-approved dyes of indocyanine green (ICG) and methylene blue (MB) without any additional excipients for cancer fluorescence imaging and combinational PTT/PDT. ICG-MB was found to exhibit good dispersion in the aqueous phase and improve the photostability and cellular uptake of free ICG and MB, thus exhibiting enhanced photothermal conversion and singlet oxygen (1O2) generation abilities to robustly ablate cancer cells under 808 nm and 670 nm laser irradiation. After intravenous injection, ICG-MB effectively accumulated at tumor sites with a near-infrared (NIR) fluorescence signal, which helped to delineate the targeted area for NIR laser-triggered phototoxicity. As a consequence, ICG-MB displayed a combinational PTT/PDT effect to potently inhibit tumor growth without causing any system toxicities in vivo. In conclusion, this minimalist, effective and biocompatible nanotheranostic would provide a promising candidate for cancer phototherapy based on current available dyes in clinic.

Homocysteine-Induced Disturbances in DNA Methylation Contribute to Development of Stress-Associated Cognitive Decline in Rats.

Chronic stress is generally accepted as the main risk factor in the development of cognitive decline; however, the underlying mechanisms remain unclear. Previous data have demonstrated that the levels of homocysteine (Hcy) are significantly elevated in the plasma of stressed animals, which suggests that Hcy is associated with stress and cognitive decline. To test this hypothesis, we analyzed the cognitive function, plasma concentrations of Hcy, and brain-derived neurotropic factor (BDNF) levels in rats undergoing chronic unpredicted mild stress (CUMS). The results showed that decreased cognitive behavioral performance and decreased BDNF transcription and protein expression were correlated with hyperhomocysteinemia (HHcy) levels in stressed rats. Diet-induced HHcy mimicked the cognitive decline and BDNF downregulation in the same manner as CUMS, while Hcy reduction (by means of vitamin B complex supplements) alleviated the cognitive deficits and BDNF reduction in CUMS rats. Furthermore, we also found that both stress and HHcy disturbed the DNA methylation process in the brain and induced DNA hypermethylation in the BDNF promoter. In contrast, control of Hcy blocked BDNF promoter methylation and upregulated BDNF levels in the brain. These results imply the possibility of a causal role of Hcy in stress-induced cognitive decline. We also used ten-eleven translocation (TET1), an enzyme that induces DNA demethylation, to verify the involvement of Hcy and DNA methylation in the regulation of BDNF expression and the development of stress-related cognitive decline. The data showed that TET1-expressing viral injection into the hippocampus inhibited BDNF promoter methylation and significantly mitigated the cognitive decline in HHcy rats. Taken together, novel evidence from the present study suggests that Hcy is likely involved in chronic stress-induced BDNF reduction and related cognitive deficits. In addition, the negative side-effects of HHcy may be associated with Hcy-induced DNA hypermethylation in the BDNF promoter. The results also suggest the possibility of Hcy as a target for therapy and the potential value of vitamin B intake in preventing stress-induced cognitive decline.

Perilla Leaf Extract Attenuates Asthma Airway Inflammation by Blocking the Syk Pathway.

Perilla frutescens (L.) Britton is a classic herbal plant used widely against asthma in China. But its mechanism of beneficial effect remains undermined. In the study, the antiallergic asthma effects of Perilla leaf extract (PLE) were investigated, and the underlying mechanism was also explored. Results showed that PLE treatment significantly attenuated airway inflammation in OVA-induced asthma mice, by ameliorating lung pathological changes, inhibiting recruitment of inflammatory cells in lung tissues and bronchoalveolar lavage fluid (BALF), decreasing the production of inflammatory cytokines in the BALF, and reducing the level of immunoglobulin in serum. PLE treatment suppressed inflammatory response in antigen-induced rat basophilic leukemia 2H3 (RBL-2H3) cells as well as in OVA-induced human peripheral blood mononuclear cells (PBMCs). Furthermore, PLE markedly inhibited the expression and phosphorylation of Syk, NF-κB, PKC, and cPLA2 both in vivo and in vitro. By cotreating with inhibitors (BAY61-3606, Rottlerin, BAY11-7082, and arachidonyl trifluoromethyl ketone) in vitro, results revealed that PLE's antiallergic inflammatory effects were associated with the inhibition of Syk and its downstream signals NF-κB, PKC, and cPLA2. Collectively, the present results suggested that PLE could attenuate allergic inflammation, and its mechanism might be partly mediated through inhibiting the Syk pathway.

The role of inflammatory cytokines in anemia and gastrointestinal mucosal injury induced by foot electric stimulation.

Foot electrical stimulation (FES) has been considered as a classic stressor that can disturb homeostasis. Acute anemia was observed in the model induced by FES. The aim of this study was to explore the role of inflammatory cytokines underlying the acute anemia and gastrointestinal (GI) mucosal injury in the FES. Twenty-four male Kunming mice (20 ± 2 g) were randomly divided into control group and experimental group. The mice were placed in a footshock chamber that can generate 0.5 mA electrical impulse periodically for 0.5 h. After the process, red blood cell count, hemoglobin concentration and hematocrit, the levels of corticotropin releasing hormone (CRH) in serum and hypothalamus, and adrenocorticotropic hormone (ACTH) in serum and pituitary were detected separately. In addition, we investigated the expressions of inflammatory cytokines (IL-1, IL-6, TNF-α, iNOS, and IL-10) in the hypothalamus and duodenum by Polymerase Chain Reaction (PCR). Results showed that this FES model induced anemia, increased CRH and ACTH activity in the serum after the FES. Moreover, the expressions of IL-1ß, IL-6, TNF-α, and iNOS were significantly increased following the process, while IL-10 was not activated. These findings suggest that anemia, the inflammatory cytokines in the hypothalamus and duodenum of the mice in the model induced by FES is closely related to GI mucosal injury/bleeding. Taken together, these results underscore the importance of anemia, GI mucosal injury/bleeding and stress, future studies would be needed to translate these findings into the benefit of affected patients.

Perilla Leaf Extract (PLE) Attenuates COPD Airway Inflammation via the TLR4/Syk/PKC/NF-κB Pathway In Vivo and In Vitro.

Chronic obstructive pulmonary disease (COPD) is a complex and heterogeneous disease characterized by persistent airflow limitation but still lacking effective treatments. Perilla frutescens (L.) Britt., an important traditional medicinal plant with excellent antioxidant and anti-inflammatory properties, is widely used for the treatment of respiratory disease in China. However, its protective activity and mechanism against COPD airway inflammation have not been fully studied. Here, the anti-inflammatory effects of the PLE were investigated, and its underlying mechanisms were then elucidated. The presented results suggested a notable effect of the PLE on airway inflammation of COPD, by significantly ameliorating inflammatory cell infiltration in lung tissue, lessening leukocytes (lymphocytes, neutrophils, and macrophages) and inflammatory mediators (interleukin 4 (IL-4), IL-6, IL-17A, interferon γ (IFN-γ), and tumor necrosis factor α (TNF-α)) in the bronchoalveolar lavage fluid (BALF) of cigarette smoke (CS)/lipopolysaccharide (LPS)-induced COPD mice in vivo and inhibiting the production of inflammatory factors (nitric oxide (NO), IL-6, and TNF-α) and intracellular reactive oxygen species (ROS) in LPS-stimulated RAW264.7 cells in vitro. For further extent, PLE treatment significantly suppressed the expression and phosphorylation of TLR4, Syk, PKC, and NF-κB p65 in vivo and their mRNA in vitro. Subsequently, by co-treating with their inhibitors in vitro, its potential mechanism via TLR4/Syk/PKC/NF-κB p65 signals was disclosed. In summary, the obtained results indicated a noteworthy effective activity of the PLE on COPD inflammation, and partly, the TLR4/Syk/PKC/NF-κB p65 axis might be the potential mechanism.

Multiple Components Rapidly Screened from Perilla Leaves Attenuate Asthma Airway Inflammation by Synergistic Targeting on Syk.

BACKGROUND: Perilla frutescens (L.) Britt., a classic medicinal plant, has been demonstrated to have anti-inflammatory and anti-allergic effects in asthma. Perilla leaves extract (PLE) exerted significant therapeutic effect against allergic asthma inflammation through Syk inhibition. But the active chemical ingredients from PLE are complex and unclear, it is difficult to fully elucidate its pharmacological mechanisms. METHODS: A method was established for rapid screening and characterization of active ingredients from PLE that targeted Syk, with which three potential active ingredients were identified. By using OVA-induced allergic asthma mouse model in vivo, OVA-induced human PBMCs inflammation model and DNP-IgE/BSA-induced RBL-2H3 cells model in vitro, the effects and mechanisms of PLE and its active components were evaluated. RESULTS: Using Syk-affinity screening method, roseoside (RosS), vicenin-2 (Vic-2) and rosmarinic acid (RosA) were identified from PLE. In vitro, PLE and its ingredients showed significant inhibitory activities against Syk, with their mixture (Mix, prepared by RosS, Vic-2 and RosA in accordance with their ratio in Syk-conjugated beads bound fraction) showing a stronger inhibitory activity. RosS, Vic-2 and RosA also showed significant effects on allergic asthma, and a synergistic effect of Mix was observed. Moreover, treatment with PLE, RosS, Vic-2, RosA, and Mix significantly inhibited the expression and phosphorylation of Syk, PKC, NF-κB p65, and cPLA2 in allergic mice lung tissue and in RBL-2H3 cells. CONCLUSION: PLE may alleviate allergic airway inflammation partly through the multiple components synergistic targeting on Syk and its downstream inflammatory pathway.

Mallotus oblongifolius extracts ameliorate ischemic nerve damage by increasing endogenous neural stem cell proliferation through the Wnt/ß-catenin signaling pathway.

Mallotus oblongifolius (MO), an edible medicinal plant from Hainan in China, shows a wide range of bioactivities. The daily consumption of MO or its extracts has been observed to ameliorate ischemic nerve injury. However the mechanisms remain unclear. In this study, the effects of MO both in vitro and in vivo were investigated. The results indicated that MO improved the motor ability, neurosensory ability, balance and grasping ability of mice with ischemic injuries, induced by bilateral common carotid artery ligation (BCCAL). In addition, MO improved the morphology of neurons, resisted the loss of neurons, and enhanced the content of the nestin protein in the cerebral cortex and subgranular zone (SGZ) area. Furthermore, in the oxygen-glucose deprivation and reperfusion (OGD/R) treated cell model, MO could effectively activate the Wnt/ß-catenin signaling pathway and promote the proliferation of neural stem cells (NSCs) and increase the protein expression levels of ß-catenin and CyclinD1. Our results suggest that Mallotus oblongifolius may be used as nutraceuticals or functional foods to alleviate ischemic nerve damage and promote recovery from ischemic stroke.

[The effect of baicalin on human brain microvascular endothelial cell under the glucose deprivation combined with hypoxia condition].

OBJECTIVE: To investigate the mechanisms of baicalin on anti-cerebral ischemic through observing the effect of baicalin on human brain microvascular endothelial cell under the glucose deprivation combined with hypoxia condition. METHODS: Human brain microvascular endothelial cells (HBMVECs) cultured in vitro were divided into the following groups: normal group, model group, baicalin high dose group, baicalin middle dose group, baicalin low dose group, nimodipine group. The kits were used to detect the cell viability, leakage rate of lactate dehydrogenase (LDH), Na(+)-K(+)-ATPase, Ca(2+)-ATPase, the concentration of Ca2+ in each group, and apoptosis rates of each group were analyzed by flow cytometry (FCM). RESULTS: Compared with the normal group, the cell viability, Na(+)-K(+)-ATPase and Ca(2+)-ATPase in model group decreased significantly (P < 0.01, P < 0.05). But the leakage rate of LDH, Ca2+ in cells and apoptosis rates increased remarkably (P < 0.01). Compared with the model group, the cell viability, Na(+)-K(+)-ATPase and Ca(2+)-ATPase increased obviously (P < 0.01, P < 0.05) in baicalin high dose group. But the leakage rate of LDH and Ca2+ in cells in baicalin high dose group decreased significantly comparing with that of model group (P < 0.05, P < 0.01). And the reduction of intracellular Ca2+ was superior to that of nimodipine group. Meanwhile, the apoptosis rates decreased significantly in both baicalin high and middle dose groups (P < 0.01). CONCLUSION: Baicalin could improve the cell viability of HBMVECs under the glucose deprivation combined with hypoxia condition. And the mechanisms were related with improving the energy metabolism, inhibiting intracellular calcium overload and decreasing the apoptosis rate of cells further.

The complete genome sequence of Natrinema sp. J7-2, a haloarchaeon capable of growth on synthetic media without amino acid supplements.

Natrinema sp. J7-2 is an extreme haloarchaeon capable of growing on synthetic media without amino acid supplements. Here we report the complete genome sequence of Natrinema sp. J7-2 which is composed of a 3,697,626-bp chromosome and a 95,989-bp plasmid pJ7-I. This is the first complete genome sequence of a member of the genus Natrinema. We demonstrate that Natrinema sp. J7-2 can use gluconate, glycerol, or acetate as the sole carbon source and that its genome encodes complete metabolic pathways for assimilating these substrates. The biosynthetic pathways for all 20 amino acids have been reconstructed, and we discuss a possible evolutionary relationship between the haloarchaeal arginine synthetic pathway and the bacterial lysine synthetic pathway. The genome harbors the genes for assimilation of ammonium and nitrite, but not nitrate, and has a denitrification pathway to reduce nitrite to N(2)O. Comparative genomic analysis suggests that most sequenced haloarchaea employ the TrkAH system, rather than the Kdp system, to actively uptake potassium. The genomic analysis also reveals that one of the three CRISPR loci in the Natrinema sp. J7-2 chromosome is located in an integrative genetic element and is probably propagated via horizontal gene transfer (HGT). Finally, our phylogenetic analysis of haloarchaeal genomes provides clues about evolutionary relationships of haloarchaea.

[Application of network pharmacology and high through-put technology on active compounds screening from traditional Chinese medicine].

OBJECTIVE: To solve the difficult in the active compounds screening from traditional Chinese medicines (TCM). METHOD: According to preliminary lab results and related literature, systematic analysis of the high-throughput technology was made. RESULT: Technologies of rapid preparation, high-throughput screening and biochip, together with the thought of drug target network, will contribute to TCM research on active ingredients screening, toxic components exclusion and molecular mechanisms. They are key technologies and ideas for modernization of TCM and research of TCM network pharmacology. CONCLUSION: High throughput technology and network pharmacology-related technologies will provide new ideas and key methods for active compounds screening from TCM.

[Study about target-network of anti-cerebral infarction neuropathy based on theory of neurovascular unit and network pharmacology].

OBJECTIVE: Potention drug-targets on anti-neuropathy of stroke were summarized, and it will provide materials for developing innovation components traditional Chinese medicine on anti-cerebral infarction neuropathy. METHOD: This article had done a series of researching work about neurovascular unit which includes three kinds of cells: neuron, gliacyte,brain microvascular endothelial cell, then signal mechanism of cell death or apoptosis of each section of stroke neuropathy was analysised by the historical documents. RESULT: There are five important pathways: inflammatory factor-MMPs pathway- Caspases, Ca2+ -mitochondrial pathway-Caspases, Ca2+ -Phospholipase-PI-3K/AK pathway, Ca2+ -radical-MAPK pathway, Ca2+ -NO-protease pathway, among all the nodes, Caspases, Ca2+, NO were the most important ones. CONCLUSION: Developing the multi-mechanism and multilevel of traditional chinese medicine under the guidance of the theories of network pharmacology and neurovascular unit will play an important role in studying the key links of signal-network of stroke neuropathy.

[Effect of acupuncture therapy on appetite of obesity patients].

OBJECTIVE: To observe the effect of acupuncture intervention on the appetite of obesity patients. METHODS: A total of 118 obesity patients were randomized into acupuncture group (76 cases, treated by true acupuncture needles) and placebo group (42 cases, treated by placebo acupuncture needles) using single-blind method. All the patients of the two groups were ordered to control their diet during the treatment. The acupoints around the umbilicus [Zhongwan (CV 12), Zhongji (CV 3), Daheng (SP 15), Xiawan (CV 10), Shimen (CV 5) and Tianshu (ST 25), etc.] and Liangqiu (ST 34), Zusanli (ST 36), and Yin-lingquan (SP 9) were punctured with filiform needles which were manipulated with uniform reducing and reinforcing method for a while tijl "Deqi" and retained for 30 min. The treatment was conducted once every other day, 12 times altogether. Body mass index (BMI), and visual analogue scale (VAS) scores of eating-desire and hunger feeling and prospective food consumption were measured before and after the treatment. The gastric fluid survival rate (GFSR) was evaluated by using ultrasound scanning. RESULTS: The BMI in the acupuncture group was obviously declined after the treatment in comparison with the placebo group (P < 0.01). Compared to the placebo group, the VAS scores of eating-desire, hunger feeling and prospective food consumption were significantly decreased in the acupuncture group ( P < 0.05), but there are no significant difference between two groups in the VAS score of gastric fullness feeling (P > 0.05). The GFSR was obviously increased in the acupuncture group compared to the placebo group (P < 0.05). CONCLUSION: Acupuncture therapy can significantly decrease BMI and delay the digesting time and control the appetite in obesity patients, which may contribute to its effect in body weight reduction.

[Clinical observation on simple obesity treated by acupuncture].

OBJECTIVE: To evaluate the effectiveness and safety of simple obesity treated by acupuncture. METHODS: By randomized single-blind clinical trial, one hundred and eighteen cases of simple obesity were divided into an acupuncture group (76 cases) and a placebo-acupuncture control group (42 cases), additionally, health control group (30 cases) was included. In acupuncture group and placebo-acupuncture control group, all the patients received a restricted diet; Zhongwan (CV 12) and Zhongji (CV 3) etc. at abdomen and Liangqiu (ST 34) and Zusanli (ST 36) etc. at limbs were selected; body mass index (BMI), Serum Total Cholesterol, triglyceride (TG), Glucose, Creatinine, urea nitrogen (BUN), Uric Acid and adverse reactions scores were observed. RESULTS: After treatment the BMI in acupuncture grown was lower than that in placebo-acupuncture control group (P < 0.01). In metabolism indices, the serum Total Cholesterol and Glucose after treatment were reduced obviously than those before treatment in acupuncture group (all P < 0.01), and there was no significant differences in other metabolism indices (all P > 0.05) in two groups. After treatment, in adverse reactions scores, the hunger sensation scores in acupuncture group was reduced than that in placebo-acupuncture control group (P < 0.05), and there was no significant differences in other indices (all P > 0.05). CONCLUSION: BMI of simple obesity was reduced by acupuncture, and the Serum Total Cholesterol and Glucose were reduced accordingly. The adverse reac tions such as weakness, nervosa and diarrhea, etc. doesn't appear after acupuncture treatment. Acupuncture therapy is one of the safe and effective methods for simple obesity.

[Effect of acupoint application therapy on expression of IL-4 and IFN-gamma in lung tissue of asthma rats].

OBJECTIVE: To observe the effect of acupoint application of Chinese herbal medicines on pathological changes of pulmonary tissue in asthma rats so as to explore its underlying mechanism. METHODS: A total of 33 Wistar rats were randomized into normal control (n=6), model (n=9), dexamethasone (DX, n=9) and acupoint-application (A-A, n=9) groups. Asthma model was established by intraperitoneal injection of 10% ovalbumin(OVA, 1 mL)and forced inhalation of atomized 2% OVA (25 mL) for 40 min, once daily for 2 weeks. "Dazhui" (GV 14). "Pishu" (BL 20), "Feishu" (BL 13) and "Shenshu" (BL 23) were selected for external application of Chinese medicinal herbs (pricklyash peel, white mustard seeds, asarum herb, etc.) for rats of A-A group, and that of control drugs (ang-kak, black rice, ginger juice, etc) for rats of DX group. Infiltration degrees of eosinophils (Eos), lymphocytes (L) and macrophages (MO) in the pulmonary tissue were observed under microscope. IL-4 and IFN-gamma expression was displayed by using immunohistochemical method. RESULTS: In comparison with control group, the infiltration degrees of E(OS). L and Mphi increased significantly in model, DX and acupoint-application groups (P < 0.05, 0.01); while compared with model group, the infiltration of E(OS), L and Mphi in DX group, E(OS) in A-A group decreased significantly (P < 0.05, 0.01). No significant differences were found between DX and A-A groups in these 3 indexes (P > 0.05). Results of immunohisto chemical staining in the lung tissue indicated that compared with control group, IL-4 expression in model increased significantly (P < 0.01), IFN-gamma expression decreased considerably in model and DX group (P < 0.01); when compared with model group, IL-4 in both A-A group and DX group decreased significantly, and IFN-gamma expression in A-A group increased considerably (P < 0.05, 0.01). No significant differences were found between DX and A-A groups in IL-4 expression (P > 0.05). CONCLUSION: Acupoint application of Chinese herbal medicines can balance Thl/Th2 and effectively reduce the infiltration of eosinophils, which may contribute to its therapeutic effect in relieving asthma.