Rapid eye movement (REM) sleep is thought to be a cortical state closer to wakefulness than non-REM sleep, yet a few brain regions regulate both states. This study identifies the glutamatergic neurons in the lateral periaqueductal gray (LPAG) that are specifically activated during wakefulness and REM sleep, as demonstrated by fiber photometry and optic tetrode recordings. Chemogenetic inhibition of LPAG vesicular glutamate transporter 2 (Vglut2) neurons reduced REM sleep and wakefulness while boosting non-REM sleep; their activation induced wakefulness, with high theta power and immobility. Chemogenetic suppression of the LPAG-sublaterodorsal tegmental nucleus projection curtailed REM sleep, and its optogenetic activation triggered REM sleep, proving that this pathway drives REM sleep. Meanwhile, LPAGVglut2 neurons promoted wakefulness through outputs to the locus coeruleus and ventral gigantocellular reticular nucleus, as supported by optogenetic manipulations. Overall, the results show that LPAGVglut2 circuits govern both REM sleep and wakefulness, uncovering a crucial region involved in the regulation of both REM sleep and wakefulness.

Innate escape behaviors, while not requiring prior learning, are shaped by an animal's learned experiences, such as previous exposure. Here, we found that learned threat experience in mice enhances flight behaviors, which is linked to increased activation of cholecystokinin-expressing neurons in the dorsal premammillary nucleus (PMdCCK neurons), a population that controls circa-strike escape responses. This heightened activity coincides with reduced inhibition from parvalbumin-expressing GABAergic neurons in the ventral tegmental nucleus of Gudden (VTgPV), which typically suppress PMdCCK activity and escape behaviors. Furthermore, threat memory prompts a prefrontal projection to stimulate the release of endocannabinoids, inhibiting the axon terminals of VTgPV neurons. The necessity of this endocannabinoid-mediated disinhibition for the observed enhancement in flight behaviors is confirmed through genetic deletion or pharmacological blockade of endocannabinoid receptors on VTgPV neurons. Thus, our study uncovers a neural mechanism by which experience amplifies innate escape behaviors, highlighting the crucial role of endocannabinoids.

OBJECTIVE:The lateral periaqueductal gray (LPAG), which mainly contains glutamatergic neurons, plays an important role in social responses, pain, and offensive and defensive behaviors. Currently, the whole-brain monosynaptic inputs to LPAG glutamatergic neurons are unknown. This study aims to explore the structural framework of the underlying neural mechanisms of LPAG glutamatergic neurons.METHODS:This study used retrograde tracing systems based on the rabies virus, Cre-LoxP technology, and immunofluorescence analysis.RESULTS:We found that 59 nuclei projected monosynaptic inputs to the LPAG glutamatergic neurons. In addition, seven hypothalamic nuclei, namely the lateral hypothalamic area (LH), lateral preoptic area (LPO), substantia innominata (SI), medial preoptic area, ventral pallidum, posterior hypothalamic area, and lateral globus pallidus, projected most densely to the LPAG glutamatergic neurons. Notably, we discovered through further immunofluorescence analysis that the inputs to the LPAG glutamatergic neurons were colocalized with several markers related to important neurological functions associated with physiological behaviors.CONCLUSION:The LPAG glutamatergic neurons received dense projections from the hypothalamus, especially nuclei such as LH, LPO, and SI. The input neurons were colocalized with several markers of physiological behaviors, which show the pivotal role of glutamatergic neurons in the physiological behaviors regulation by LPAG.

Surface engineering of nanoparticles has been widely used in biosensing and assays, where sensitivity was mainly limited by plasmonic colour change or electrochemical responses. Here, we report a novel biomimetic sensing strategy involving protein-modified gold nanoparticles (AuNPs), where the modulation strategy was inspired by gastropods in inhibition of coffee-ring effects in their trail-followings. The so-called coffee-ring effect presents the molecular behaviour of AuNPs to a macroscopic ring through aggregation, and thus greatly improves sensitivity. The assay relies upon the different assembly patterns of AuNPs against analytes, resulting in the formation or suppression of coffee-ring effects by the different surface engineering of AuNPs by proteins and peptides. The mechanism of the coffee-ring formation process is examined through experimental characterizations and computational simulations. A practical coffee-ring effect assay is developed for a proof-of-concept target, amyloid β (1–42), which is a typical biomarker of Alzheimer’s disease. A novel quasi-titrimetric protocol is constructed for quantitative determination of the target molecule. The assay shows excellent selectivity and sensitivity for the amyloid β monomer, with a low detection limit of 20 pM. Combined with a fluorescent staining technique, the assay is designed as a smart sensor for amyloid β detection and fibrillation evaluation in rat cerebrospinal fluids, which is a potential point-of-care test for Alzheimer’s disease. Connections between amyloid fibrillation and different courses of brain ischaemia are also studied, with improved sensitivity, lower sample volumes that are required, convenience for rapid detection, and point-of-care testing.

Good sleep quality is essential for maintaining the body’s attention during wakefulness, which is easily affected by external factors such as an ambient temperature. However, the mechanism by which an ambient temperature influences sleep–wake behaviors remains unclear. The dorsomedial hypothalamus (DMH) has been reported to be involved in thermoregulation. It also receives projection from the preoptic area, which is an important region for sleep and energy homeostasis and the suprachiasmatic nucleus—a main control area of the clock rhythm. Therefore, we hypothesized that the DMH plays an important role in the regulation of sleep related to ambient temperatures. In this study, we found that cold exposure (24/20/16/12 °C) increased wakefulness and decreased non–rapid eye movement (NREM) sleep, while warm exposure (32/36/40/44 °C) increased NREM sleep and decreased wakefulness compared to 28 °C conditions in wild-type mice. Then, using non-specific and specific apoptosis, we found that lesions of whole DMH neurons and DMH γ–aminobutyric acid (GABA)-ergic neurons induced by caspase-3 virus aggravated the fluctuation of core body temperature after warm exposure and attenuated the change in sleep–wake behaviors during cold and warm exposure. However, chemogenetic activation or inhibition of DMH GABAergic neurons did not affect the sleep–wake cycle. Collectively, our findings reveal an essential role of DMH GABAergic neurons in the regulation of sleep–wake behaviors elicited by a change in ambient temperature.

Sleep and wakefulness are basic behavioral states that require coordination between several brain regions, and they involve multiple neurochemical systems, including neuropeptides. Neuropeptides are a group of peptides produced by neurons and neuroendocrine cells of the central nervous system. Like traditional neurotransmitters, neuropeptides can bind to specific surface receptors and subsequently regulate neuronal activities. For example, orexin is a crucial component for the maintenance of wakefulness and the suppression of rapid eye movement (REM) sleep. In addition to orexin, melanin-concentrating hormone, and galanin may promote REM sleep. These results suggest that neuropeptides play an important role in sleep–wake regulation. These neuropeptides can be divided into three categories according to their effects on sleep–wake behaviors in rodents and humans. (i) Galanin, melanin-concentrating hormone, and vasoactive intestinal polypeptide are sleep-promoting peptides. It is also noticeable that vasoactive intestinal polypeptide particularly increases REM sleep. (ii) Orexin and neuropeptide S have been shown to induce wakefulness. (iii) Neuropeptide Y and substance P may have a bidirectional function as they can produce both arousal and sleep-inducing effects. This review will introduce the distribution of various neuropeptides in the brain and summarize the roles of different neuropeptides in sleep–wake regulation. We aim to lay the foundation for future studies to uncover the mechanisms that underlie the initiation, maintenance, and end of sleep–wake states.

The present study aimed to assess the anticancer cell and anticancer stem cell (CSC) effects of GANT61, and its regulatory influence on the Wnt/β‑catenin and Notch signalling pathways in colorectal cancer (CRC). HT‑29 and HCT‑116 cells were treated with 0, 2.5, 5, 10, 20 or 40 µM GANT61, after which relative cell viability and the expression of Gli1, β‑catenin and Notch1, as well as the percentage of CD133+ cells, were detected. Subsequently, HT‑29/HCT‑116 cells and CSCs were treated with 20 µM GANT61, 10 mM of the Wnt/β‑catenin pathway agonist HLY78, and 30 mM of the Notch pathway agonist JAG1 (alone or in combination), which was followed by the assessment of cell viability and apoptosis. In both cell lines, GANT61 reduced relative cell viability in a time‑ and dose‑dependent manner, inhibited Gli1, β‑catenin and Notch1 expression, and decreased the percentage of CD133+ cells in a dose‑dependent manner. Furthermore, HLY78 and JAG1 were both found to improve the relative viability, while downregulating the apoptosis of untreated and GANT61‑treated HT‑29 and HCT‑116 cells. Moreover, Wnt/β‑catenin and Notch signalling pathway activity were upregulated in CSCs isolated from HT‑29 and HCT‑116 cells, compared with the associated control groups. GANT61 also reduced the viability of HT‑29 and HCT‑116 cells and increased apoptosis, whereas HLY78 and JAG1 treatment resulted in the opposite effect. Moreover, both HLY78 and JAG1 attenuated the effects of GANT61 on cellular viability and apoptosis. In conclusion, GANT61 was found to effectively eliminate cancer cells and CSCs by blocking the Wnt/β‑catenin and Notch signalling pathways in CRC.

Quantitative drug release is important for improving therapeutic efficiency and avoiding side effects. While using long-term delivery system for repeated therapies, it is indispensable but challenging to accurately control the drug dosing. Here, a photocleavable prodrug loaded hydrogel is proposed for near infrared (NIR) light-triggered quantitative pulsed drug release. IR783, a commercially available NIR fluorescent dye, is conjugated with methyl honokiol (mHNK) to give a photocleavable IR783-mHNK prodrug. Injectable glycol chitosan (GC) hydrogel is chosen as a reservoir, in which IR783-mHNK can be efficiently loaded via electrostatic and hydrophobic interactions. Upon 680 nm light-emitting diode (LED) light irradiation, IR783-mHNK cleaves and mHNK is released. Notably, it is found that IR783-mHNK presents synchronous photocleavage-fluorescence bleaching phenomenon. The released amount of mHNK is visible by measuring the residual fluorescent intensity of hydrogel. Quantitative drug release is achieved by controlling irradiation duration and the drug release process is visible by fluorescence imaging. The prodrug-loaded hydrogel shows good stability, minimum leakage and efficient light responsibility both in vitro and in vivo. After light triggering, monitorable quantitative mHNK release and on-demand sleep-promotiing effect are verified in mice without toxicities.

Background and Purpose Modafinil is a potent eugeroic (wakefulness-promoting) drug that is prescribed to treat narcolepsy and has a low incidence of abuse. Although previous studies have shown that modafinil-induced arousal depends on the dopamine receptors and transporters, the specific part/s of the dopamine transmitter system underlying this mechanism remained unclear. Here, we investigated the role of mesencephalic dopamine neurons in modafinil-evoked arousal. Experimental Approach A dopamine indicator (dLight1.1) was employed to detect dopamine changes in the nucleus accumbens (NAc) and dorsal striatum. We specifically lesioned mesencephalic dopamine neurons via diphtheria toxin (DTA) in the dopamine transporter (DAT)-Cre mice. Then, the sleep-wake states were recorded to evaluate the effect of modafinil on arousal. Finally, the extent of DTA-induced lesions was determined by immunohistochemistry. Key Results Modafinil promptly increased dopamine levels in the NAc and dorsal striatum in a dose-dependent manner. Lesioning of dopamine neurons in the substantia nigra pars compacta (SNc) or ventral tegmental area (VTA) had no significant effects on physiological sleep-wake cycles. Modafinil at 90 mg center dot kg(-1) increased continuous wakefulness for 355 min in control mice. However, these effects were slightly decreased by 6.7% in the SNc-lesioned mice and were prominently diminished by 32.8% in VTA-lesioned mice. Furthermore, the modafinil-induced arousal was completely blocked in the SNc-VTA-lesioned mice, whereas lesions of the dorsal raphe nucleus had no effect. Conclusion and Implications Taken together, our findings indicate that mesencephalic dopamine neurons are essential for modafinil-induced arousal.

As one of the fundamental sleep states, rapid eye movement (REM) sleep is believed to be associated with dreaming and is characterized by low-voltage, fast electroencephalographic activity and loss of muscle tone. However, the mechanisms of REM sleep generation have remained unclear despite decades of research. Several models of REM sleep have been established, including a reciprocal interaction model, limit-cycle model, flip-flop model, and a model involving gamma-aminobutyric acid, glutamate, and aminergic/orexin/melanin-concentrating hormone neurons. In the present review, we discuss these models and summarize two typical disorders related to REM sleep, namely REM sleep behavior disorder and narcolepsy. REM sleep behavior disorder is a sleep muscle-tone-related disorder and can be treated with clonazepam and melatonin. Narcolepsy, with core symptoms of excessive daytime sleepiness and cataplexy, is strongly connected with orexin in early adulthood.

Depression, rapid eye movement (REM) sleep behavior disorder, and altered olfaction are often present in Parkinson's disease. Our previous studies demonstrated the role of the olfactory bulb (OB) in causing REM sleep disturbances in depression. Furthermore, adenosine A2A receptors (A2AR) which are richly expressed in the OB, play an important role in the regulation of REM sleep. Caffeine, an adenosine A1 receptors and A2AR antagonist, and other A2AR antagonists were reported to improve olfactory function and restore age-related olfactory deficits. Therefore, we hypothesized that the A2AR neurons in the OB may regulate olfaction or odor-guided behaviors in mice. In the present study, we employed chemogenetics to specifically activate or inhibit neuronal activity. Then, buried food test and olfactory habituation/dishabituation test were performed to measure the changes in the mice's olfactory ability. We demonstrated that activation of OB neurons or OB A2AR neurons shortened the latency of buried food test and enhanced olfactory habituation to the same odors and dishabituation to different odors; inhibition of these neurons showed the opposite effects. Photostimulation of ChR2-expressing OB A2AR neuron terminals evoked inward current in the olfactory tubercle (OT) and the piriform cortex (Pir), which was blocked by glutamate receptor antagonists 2-amino-5-phosphonopentanoic acid and 6-cyano-7nitroquinoxaline-2,3-dione. Collectively, these results suggest that the OB mediates olfaction via A2AR neurons in mice. Moreover, the excitatory glutamatergic release from OB neurons to the OT and the Pir were found responsible for the olfaction-mediated effects of OB A2AR neurons.

Non-specific immune responses to antigens have been demonstrated as being enhanced during chronic hepatitis B virus (HBV) infection. Here, we evaluated the role of interleukin-10 (IL-10)-producing regulatory B-cells (Bregs) in the pathogenesis of HBV-related liver fibrosis (HBV-LF) and assessed their immunoregulatory effects. Sixty-seven patients diagnosed with chronic hepatitis B (CHB) were enrolled in this study. Numbers and frequencies of peripheral B-cells (memory CD19+CD24hiCD27+ cells, immature/transitional CD19+CD24hiCD38hi cells, mature CD19+CD24intCD38int cells) were tested and analysed. Flow cytometry-sorted CD4+T cells were cultured with autologous Bregs to elucidate the effects of Bregs on CD4+T cells, including effector T and regulatory T-cells (Tregs). The potential immunoregulatory mechanism of Bregs was also investigated. The numbers of total B-cells and Bregs were enriched in CHB patients. The frequency of Bregs was negatively correlated with elevated alanine aminotransferase (ALT) and histological inflammation grades (G), but positively correlated with advanced histological fibrosis stages (S) and enhanced HBV replication. The phenotype of Bregs was predominantly characterized as CD19+CD24hiCD38hi. In co-culture with Bregs, CD4+CD25−T cells from CHB patients produced less interferon-γ (IFN-γ) and IL-17 but more IL-4 than CD4+CD25−T cells alone, whereas their conversions into Tregs and IL-10+T cells were enhanced. In addition, Breg depletion in CHB samples dramatically decreased Treg numbers and expression of cytotoxic T-lymphocyte associated antigen-4 (CTLA-4), IL-10 and transforming growth factor-β (TGF-β). Moreover, the observed regulatory effect was partly dependent on IL-10 release and cell-to-cell contact. Elevated Bregs can suppress effector T but enhance Treg functions, which might influence immune tolerance in chronic HBV infection.

OBJECTIVE This study aimed to determine whether the human immunodeficiency virus (HIV) exists in giant idiopathic esophageal ulcers in the patients with acquired immune deficiency syndrome (AIDS). METHODS 16 AIDS patients with a primary complaint of epigastric discomfort were examined by gastroscopy. Multiple and giant esophageal ulcers were biopsied and analyzed with pathology staining and reverse transcription-polymerase chain reaction (RT-PCR) to determine the potential pathogenic microorganisms, including HIV, cytomegalovirus (CMV) and herpes simplex viruses (HSV). RESULTS HIV was detected in ulcer samples from 12 out of these 16 patients. Ulcers in 2 patients were infected with CMV and ulcers in another 2 patients were found HSV positive. No obvious cancerous pathological changes were found in these multiple giant esophageal ulcer specimens. CONCLUSION HIV may be one of the major causative agents of multiple benign giant esophageal ulcers in AIDS patients.

Objective To develop experimental retinal vein occlusion with photodynamic method in combination with rose bengal dye in SD rats, and investigate the morphologic and histologic changes in the retina. Methods After SD rats tail intravenous injection of rose bengal,the rats retinal veins near the disc of 1-3 PD were exposed to diode laser(wave-length 532 nm) and formed complete vascular occlusion and the 1 PD thrombus could be seen. The eyes were observed with eye fundus check and fundus fluorescein angiography(FFA) every other day for three weeks. Then the eyes were enucleated and prepared for light and electron microscope examination. Results Retinal vein occlusion was developed successfully by combining photodynamic thrombosis and rose bengal dye in SD rats.Histopathologic features were consistent with changes of retinal vein occlusion. Furthermore,formation of retinal vein thrombus was reliable. Conclusions Photodynamic method combined with rose bengal dye is a simple,reliable and definitive experimental technique to produce rats retinal vein occlusion.

Data of 14 patients(23 eyes) with central serous choroidoretinaopathy(CSC) or bullous retinal detachment induced by steroids application were analyzed retrospectively.Deterioration of visual acuity(VA) and metamorphosis were major symptoms.Object findings were located at the posterior pole,which included one or two serous translucent shallow retinal detachment(RD);one or several subretinal yellowish exudates some accompanied with inferior shallow RD;bullous RD with subretinalfibrinous exudates.Fluorescein fundus angiography showed one or several retinal pigment epithelium leakage in all cases.Optical coherence tomography found sensory RD with subretinal hyper-reflecting materials in most cases.All these findings were in accordance with the diagnosis of CSC.Cessation of steroids was undertaken.RD gradually reattached within 2~10 months.Among them,cases with serous RD or subretinal yellowish exudates recovered well with final VA ≥0.5.The worst VA was found in bullous RD patients with VA <0.1.The paper discussed how to distinquish whether inadequate dosage of steroids or steroids caused worse of VA in steroids induced CSC.

Objective: Methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms have been reported to be associated with pancreatic cancer, but the published studies had yielded inconsistent results.We therefore performed the present meta-analysis. Methods: A search of Google scholar, PubMed, Cochrane Library and CNKI databases before April 2012 was conducted to summarize associations of MTHFR polymorphisms with pancreatic cancer risk. Assessment was with odds ratios (ORs) and 95% confidence intervals (CIs). Publication bias were also calculated. Results: Four relative studies on MTHFR gene polymorphisms (C667T and A1298C) were involved in this meta-analysis. Overall, C667T(TT vs. CC : OR = 1.61, 95%CI = 0.78 - 3.34; TT vs. CT : OR = 1.41, 95%CI = 0.88-2.25; dominant model: OR = 0.68, 95%CI = 0.40-1.17; recessive model: OR = 0.82, 95%CI = 0.52-1.30) and A1298C(CC vs. AA:OR=1.01, 95%CI=0.47-2.17; CC vs. AC: OR=0.99,95%CI=0.46-2.14; dominant model: OR=1.01, 95%CI = 0.47-2.20; recessive model: OR = 1.01, 95%CI = 0.80-1.26) did not increase pancreatic cancer risk. Conclusion: This meta-analysis indicated that MTHFR polymorphisms (C667T and A1298C) were not associated with pancreatic cancer risk.

To investigate anti-infective treatments in HIV-infected surgical patients during the perioperative period.

Background Surgical site infection (SSI) are the third most frequently reported nosocomial infection, and the most common on surgical wards. HIV-infected patients may increase the possibility of developing SSI after surgery. There are few reported date on incidence and the preventive measures of SSI in HIV-infected patients. This study was to determine the incidence and the associated risk factors for SSI in HIV-infected patients. And we also explored the preventive measures. Methods A retrospective study of SSI was conducted in 242 HIV-infected patients including 17 patients who combined with hemophilia from October 2008 to September 2011 in Shanghai Public Health Clinical Center. SSI were classified according to Centers for Disease Control and Prevention (CDC) criteria and identified by bedside surveillance and post-discharge follow-up. Data were analyzed using SPSS 16.0 statistical software (SPSS Inc., Chicago, IL). Results The SSI incidence rate was 47.5% (115 of 242); 38.4% incisional SSIs, 5.4% deep incisional SSIs and 3.7% organ/space SSIs. The SSI incidence rate was 37.9% in HIV-infected patients undergoing abdominal operation. Patients undergoing abdominal surgery with lower preoperative CD4 counts were more likely to develop SSIs. The incidence increased from 2.6% in clean wounds to 100% in dirty wounds. In the HIV-infected patients combined with hemophilia, the mean preoperative albumin and postoperative hemoglobin were found significantly lower than those in no-SSIs group (P<0.05). Conclusions SSI is frequent in HIV-infected patients. And suitable perioperative management may decrease the SSIs incidence rate of HIV-infected patients.

Objective:To investigate the molecular mechanism by which triamcinolone acetonide(TA ) may reduce edema in a SD rat model of retinal vein occlusion(RVO).Methods:After baseline ophthalmoscopic examination and fundus photography, a RVO was created photothrombotically in both eyes of 32 rats,using green photocoagulation and intravenous Rose Bengal. Following this,32 eyes were injected intravitreally with 320μg/8μl TA.After several days,all eyes were re-examined.Fluorescein angiography,in addition to ophthalmoscopy and fundus photography,was performed.Following sacrifice of the animals, the eyes were enucleated and processed,vascular endothelial growth factor(VEGF) was detected by ELISA and fluorescent immunohistochemistry,and compared and analyzed it by light microscope and image analysis software.Results:The changes of eye fundus amelioration were observed obviously in experimental group.Retinal VEGF levels were significantly lower in the TA-treated eyes as compared with the untreated eyes(P0.01).Conclusions:Intravitreal TA can down-regulate VEGF. This event may contribute to a reduction in the blood retinal barrier breakdown that occurs in RVO.

To investigate the management and prognosis of HIV infected patients with postoperative sepsis, the clinical data of 126 HIV-infected patients aged 20 to 74 years including 114 males and 12 females were retrospectively analyzed. Patients were divided into 3 groups according to the pre-operative level of CD4+ cells: Group A (n = 73), <200 cells/μl; Group B (n = 36): 200 cells/μl ≤ CD4 + cells < 350 cells/μl; Group C (n = 17): ≥350 cells/μl. The operation was carefully performed aiming to reduce the damage to tissues. Antibiotic prophylaxis, treatments with anti-tuberculous, antifugal and anti-retroviral drugs and timely treatment of complications were carried out. In Group A, B and C, postoperative sepsis occurred in 32, 8 and 3 patients, respectively, of whom 4, 1 and 0 died of multiple organ dysfunction syndrome (MODS), respectively. Analysis showed the proportion of patients with sepsis in Group A was significantly higher than that in Group B and Group C, but no significant difference was found between Group B and C. The HIV infected patients with CD4 + cells <200 cells/μl had significantly increased risk for postoperative sepsis. Appropriate peri-operative treatment for HIV-infected patients may achieve favorable outcome. Key words: Acquired immune deficiency syndrome, operative-risk evaluation,complication, sepsis.