In situ Injection of pH- and Temperature-Sensitive Nanomaterials Increases Chemo-Photothermal Efficacy by Alleviating the Tumor Immunosuppressive Microenvironment.

Purpose: Triple-negative breast cancer (TNBC) is challenging to treat with traditional "standard of care" therapy due to the lack of targetable biomarkers and rapid progression to distant metastasis. Methods: We synthesized a novel combination regimen that included chemotherapy and photothermal therapy (PTT) to address this problem. Here, we tested a magnetic nanosystem (MNs-PEG/IR780-DOX micelles) loaded with the near-infrared (NIR) photothermal agent IR780 and doxorubicin (DOX) to achieve chemo-photothermal and boost antitumor immunity. Intraductal (i.duc) administration of MNs-PEG/IR780-DOX could increase the concentration of the drug in the tumor while reducing systemic side effects. Results: We showed more uptake of MNs-PEG/IR780-DOX by 4T1-luc cells and higher penetration in the tumor. MNs-PEG/IR780-DOX exhibited excellent photothermal conversion in vivo and in vitro. The release of DOX from MNs-PEG/IR780-DOX is pH- and temperature-sensitive. Facilitated by i.duc administration, MNs-PEG/IR780-DOX displayed antitumor effects and prevented distant organs metastasis under NIR laser (L) irradiation and magnetic field (MF)while avoiding DOX-induced toxicity. More importantly, MNs-PEG/IR780-DOX alleviated tumor immunosuppressive microenvironment by increasing tumor CD8+ T cells infiltration and reducing the proportion of myeloid-derived suppressor cells (MDSCs) and Tregs. Conclusion: Intraductal administration of pH- and temperature-sensitive MNs-PEG/IR780-DOX with L and MF had the potential for achieving minimally invasive, targeted, and accurate treatment of TNBC.

Serum containing Gengnianchun formula suppresses amyloid ß­induced inflammatory cytokines in BV­2 microglial cells by inhibiting the NF­κB and JNK signaling pathways.

As the resident macrophages of the brain's innate immune system, microglial cells are key modulators in the neurodegenerative disease Alzheimer's disease (AD). In particular, the activation and accumulation of microglial cells around amyloid plaques is considered to result in chronic neuroinflammation. Although the pathologic mechanism remains to be fully elucidated, inflammation has been shown to be critical in the pathogenesis of AD. The Gengnianchun (GNC) formula has long been used to treat perimenopausal syndrome clinically, and is particularly effective in improving learning ability and memory. Our previous study demonstrated that GNC formula had an anti­inflammatory effect and offered neuroprotection in animal experiments. In the present study, the anti­inflammatory properties of GNC and its underlying mechanism of action were examined in BV­2 microglial cells. Amyloid­ß peptide (Aß)­stimulated microglial cells were examined for the production of proinflammatory cytokines and the underlying signaling pathways. Compared with the normal control group, the protein expression levels of IL­1ß and TNF­α were significantly increased following treatment with Aß (P<0.01), but medicated rat serum containing GNC formula (MRS) could significantly attenuated the Aß­induced secretion of these pro­inflammatory cytokines. It was identified by CCK­8 assay that the viability of the BV­2 cells was not reduced following treatment with various concentrations of MRS. The phosphorylation of factor­κB (NF­κB) and c­Jun N­terminal kinase (JNK) was markedly increased following treatment with Aß, compared with the normal control group (P<0.01). However, treatment with MRS resulted in a significant reduction in the phosphorylation of NF­κB (P<0.05). These results suggested that MRS suppressed the Aß­induced inflammatory response of microglial cells by inhibiting the NF­κB and JNK signaling pathways. These novel findings provide insights into the development of GNC formula as a therapeutic agent for the treatment of neurodegenerative disorders.

Effects of Gengnianchun on learning and memory ability, neurotransmitter, cytokines, and leptin in ovariectomized rats.

This study aimed to evaluate the beneficial effects of a traditional Chinese medicine named Gengnianchun (GNC) in ovariectomized Sprague-Dawley rats. The rats were randomly categorized into sham-operated group (Sham), saline-treated ovariectomized group (OVX), GNC-treated ovariectomized group (OVX+GNC), estradiol valerate-treated ovariectomized group (OVX+E). GNC and estradiol was administered for 1 month at dosages of 125 and 0.1 mg/day, respectively. Ovariectomy caused deterioration of learning and memory ability (P < 0.05), which was restored by treatment with GNC and estradiol (P < 0.05). Estrogen level and endometrial thickness significantly decreased in the OVX group (P < 0.05). These parameters significantly increased in the OVX+E group (P < 0.05) but did not change in the OVX+GNC group (P > 0.05). GNC and estradiol significantly increased the levels of norepinephrine (NE) and dopamine (DA) and decreased the levels of 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) in the hypothalamus (P < 0.05). The levels of interleukin-1 beta (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) significantly decreased and the levels of interleukin-2 (IL-2) and interferon-gamma (IFN-γ) increased in the OVX+GNC and OVX+E groups compared with those in the OVX group (P < 0.05). OVX rats treated with GNC and estradiol further exhibited reversed ovariectomy-induced weight gain and leptin resistance (P < 0.05). These results indicated that GNC demonstrated phytoestrogen-like properties without the side effects of estradiol valerate. Thus, GNC may confer protective and beneficial effects for management of menopausal syndrome.

Repeated in vivo exposure of cocaine induces long-lasting synaptic plasticity in hypocretin/orexin-producing neurons in the lateral hypothalamus in mice.

Hypocretin (orexin), a neuropeptide synthesized exclusively in the perifornical/lateral hypothalamus, is critical for drug seeking and relapse, but it is not clear how the circuitry centred on hypocretin-producing neurons (hypocretin neurons) is modified by drugs of abuse and how changes in this circuit might alter behaviours related to drug addiction. In this study, we show that repeated, but not single, in vivo cocaine administration leads to a long-lasting, experience-dependent potentiation of glutamatergic synapses on hypocretin neurons in mice following a cocaine-conditioned place preference (CPP) protocol. The synaptic potentiation occurs postsynaptically and probably involves up-regulation of AMPA-type glutamate receptors on hypocretin neurons. Phosphorylation of cAMP response element-binding protein (CREB) is also significantly increased in hypocretin neurons in cocaine-treated animals, suggesting that CREB-mediated pathways may contribute to synaptic potentiation in these cells. Furthermore, the potentiation of synaptic efficacy in hypocretin neurons persists during cocaine withdrawal, but reverses to baseline levels after prolonged abstinence. Finally, the induction of long-term potentiation (LTP) triggered by a high-frequency stimulation is facilitated in hypocretin neurons in cocaine-treated mice, suggesting that long-lasting changes in synapses onto hypocretin neurons would probably be further potentiated by other stimuli (such as concurrent environmental cues) paired with the drug. In summary, we show here that hypocretin neurons undergo experience-dependent synaptic potentiation that is distinct from that reported in other reward systems, such as the ventral tegmental area, following exposure to cocaine. These findings support the idea that the hypocretin system is important for behavioural changes associated with cocaine administration in animals and humans.

Huanglian-Jie-Du-Tang extract protects against chronic brain injury after focal cerebral ischemia via hypoxia-inducible-factor-1α-regulated vascular endothelial growth factor signaling in mice.

Huanglian-Jie-Du-Tang (HJDT) is a traditional Chinese herbal formula which is widely used clinically. In this study, we investigated the effects of an aqueous (HJDTaq) and an ethanolic (HJDTet) extract of HJDT on chronic brain injury after focal cerebral ischemia in mice. The ischemia was induced by occlusion of the right middle cerebral artery for 30 min. HJDTaq (4 g/kg) and HJDTet (200, 400, 800 mg/kg) were orally administered for 21 d from day 7 before ischemia to day 14 after ischemia. The survival rate decreased to less than 50% at 35 d after ischemia. HJDTet at 400 mg/kg increased the survival rate. HJDTaq (4 g/kg) and HJDTet (400, 800 mg/kg) significantly attenuated the neurological dysfunction, brain atrophy and infarct volume after ischemia. There were few cells positive for CD31, hypoxia-inducible-factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and Flk-1 in the sham control. After ischemia, the number increased. HJDTaq (4 g/kg) and HJDTet (400 or 800 mg/kg) further increased the numbers of CD31, HIF-1α, VEGF and Flk-1-positive cells in the ischemic hemisphere. We conclude that HJDTaq and HJDTet have neuroprotective effects on chronic brain injury after focal cerebral ischemia and lead to accelerated angiogenesis by HIF-1α-regulated VEGF signaling.

Prolonged wakefulness induces experience-dependent synaptic plasticity in mouse hypocretin/orexin neurons.

Sleep is a natural process that preserves energy, facilitates development, and restores the nervous system in higher animals. Sleep loss resulting from physiological and pathological conditions exerts tremendous pressure on neuronal circuitry responsible for sleep-wake regulation. It is not yet clear how acute and chronic sleep loss modify neuronal activities and lead to adaptive changes in animals. Here, we show that acute and chronic prolonged wakefulness in mice induced by modafinil treatment produced long-term potentiation (LTP) of glutamatergic synapses on hypocretin/orexin neurons in the lateral hypothalamus, a well-established arousal/wake-promoting center. A similar potentiation of synaptic strength at glutamatergic synapses on hypocretin/orexin neurons was also seen when mice were sleep deprived for 4 hours by gentle handling. Blockade of dopamine D1 receptors attenuated prolonged wakefulness and synaptic plasticity in these neurons, suggesting that modafinil functions through activation of the dopamine system. Also, activation of the cAMP pathway was not able to further induce LTP at glutamatergic synapses in brain slices from mice treated with modafinil. These results indicate that synaptic plasticity due to prolonged wakefulness occurs in circuits responsible for arousal and may contribute to changes in the brain and body of animals experiencing sleep loss.

[Therapeutic effect of acupuncture on allergic rhinitis and its effects on immunologic function].

OBJECTIVE: To compare therapeutic effects of acupuncture and auricular point pressing therapy on allergic rhinitis. METHODS: One hundred and fifty cases were randomly divided into an acupuncture group, an auricular point pressing group and a medication control group, 50 cases in each group. The acupuncture group were treated with acupuncture for 6 consecutive days, once each day, with an interval of one day; and the auricular point pressing group were alternatively treated twice each week; the medication group were treated with oral administration of western medicine saitezan, once daily, 10 mg once. They were treated for 4 weeks, and the short-term and long-term therapeutic effects, and effects on serum immunological indexes were compared among the three groups. RESULTS: The acupuncture group and the auricular point pressing group in the short-term therapeutic effect and improvement of cumulative scores of symptoms and signs, and the long-term total therapeutic effect and the improvement of cumulative scores of symptoms and signs were better than the medication group (P<0.01, P<0.05), with no significant differences between the acupuncture group and the auricular point pressing group. Serum IgE, IL-4 contents decreased and IFN-gamma level did not significantly change in the three groups. CONCLUSION: Acupuncture and auricular pressing therapy in improvement of symptoms and signs are better than the medication, and the therapeutic effect is carried out by inhibiting the differentiation from Th cells to Th2 cells, adjusting the imbalance of Th1/Th2 cells and decrease the synthesis of IgE to inhibit allergic rhinitis.

RP-HPLC determination of puerarin in Chinese traditional medicinal preparations containing pueraria.

Puerarin in a Gegen(Pueraria)-based Chinese traditional medicinal preparations, Ganmao Qingre Granules, from four different pharmaceutical manufacturers and in two different dosage forms, were determined using RP-HPLC with methanol-5 mmol l(-1) KH(2)PO(4) (pH = 4.0) (V:V = 27:73) as mobile phase and UV detection at 248 nm. Ultrasonication and reflux were compared as pretreatment procedures for the sample. Linear range over 0.2-200 microg ml(-1) of puerarin was obtained, and the limit of detection was 0.1 microg ml(-1). Recovery was within 99.7-103%.