The nucleus accumbens (NAc) plays an important role in various emotional and motivational behaviors that rely on heightened wakefulness. However, the neural mechanisms underlying the relationship between arousal and emotion regulation in NAc remain unclear. Here, we investigated the roles of a specific subset of inhibitory corticotropin-releasing hormone neurons in the NAc (NAc^CRH) in regulating arousal and emotional behaviors in mice. We found an increased activity of NAc^CRH neurons during wakefulness and rewarding stimulation. Activation of NAc^CRH neurons converts NREM or REM sleep to wakefulness, while inhibition of these neurons attenuates wakefulness. Remarkably, activation of NAc^CRH neurons induces a place preference response (PPR) and decreased basal anxiety level, whereas their inactivation induces a place aversion response and anxious state. NAc^CRH neurons are identified as the major NAc projection neurons to the bed nucleus of the stria terminalis (BNST). Furthermore, activation of the NAc^CRH-BNST pathway similarly induced wakefulness and positive emotional behaviors. Taken together, we identified a basal forebrain CRH pathway that promotes the arousal associated with positive affective states.
Animals ; Septal Nuclei/metabolism* ; Nucleus Accumbens/physiology* ; Corticotropin-Releasing Hormone/metabolism* ; Wakefulness/physiology* ; Neurons/metabolism* ; Male ; Mice ; Mice, Inbred C57BL ; Neural Pathways/physiology* ; Anxiety/physiopathology* ; Reward

OBJECTIVE：To investi gate the potential mechanism of couplet medicine of Cuscutae Semen-Lycii Fructus in the treatment of premature ovarian failure. METHODS ：Main active components and related targets of couplet medicine of Cuscutae Semen-Lycii Fructus in the treatment of premature ovarian failure were obtained from TCMSP ，GeneCards and OMIM database. The intersection genes between them were screened using Venn online tool. Cytoscape 3.7.0 software was adopted to establish the active ingredients-target network and the PPI network. GO and KEGG pathway enrichment analysis on intersection genes were carried out by DAVID database. Finally ，an active component-target-key pathway network was constructed. RESULTS ：Totally 42 active components ，231 and 1 913 targets for active components and disease were obtained from couplet medicine of Cuscutae Semen-Lycii Fructus. The components with high node degree included quercetin ，kaempferol，β-sitosterol，isorhamnetin，glycitein， stigmasterol and sesamin ，etc. There were 149 intersection genes between the active component targets and premature ovarian failure targets. PPI network contained 149 nodes and 2 970 edges，with an average node degree of 39.9 and an average medium of 0.005 4. The results of GO analysis showed that molecule function of the above-mentioned genes mainly involved protein binding ， enzyme binding ，etc. Biological process mainly included that positive regulatio n of transcription from RNA polymerase Ⅱ promoter，positive regulation of transcription DNA-templated ， Cell components mainly included nucleus ，cytoplasm，etc. Signaling pathway mainly involved cancer signaling pathway ， hepatitis B signling pathway ，PI3K/AKT signaling pathway ， MAPK signaling pathway ， etc. The results of active 617693370@qq.com component-target-key pathway network showed that active components of Cuscutae Semen and Lycii Fructus were flavonoids and alcohols ；key target included AKT 1，TP53， VEGFA，IL6，TNF，etc. Signaling pathway mainly involved cancer signaling pathway ，hepatitis B signaling pathway ，PI3K/AKT signaling pathway ，MAPK signaling pathway ，etc. CONCLUSIONS ：Through PI 3K/AKT signaling pathway and MAPK signaling pathway，the active components of couplet medicine of Cuscutae Semen-Lycii Fructus may act on AKT 1，TP53 and other targets ， and then play a therapeutic role on premature ovarian failure. The Potential active components stigmasterol ，sesamin and potential targets IL 6，TNF were found.

Vitamins are classified as either fat-soluble (vitamins A, D, E, K) or water-soluble (vitamins B and vitamin C). Traditional methods of immunoassay have only been developed for vitamins D,B6, B9 and B12. However, they cannot distinguish between vitamin subtypes such as D2, D3 and associated epi isomers (which has higher leveks in infants),giving false positive or negative results. Mass spectrometry has become a gold standard method for small molecule analysis in biological samples with its advantages in speed,resolution,sensitivity and specificity. It is widely used in clinical research and diagnosis and provides an efficient method for simultaneous detection of multivitamins in one injection using one low volume sample collection.

OBJECTIVE To analyze disease-related clinical features and therapeutic effects of basal cell adenoma in head and neck. METHODS Clinical data of 9 patients with pathologically diagnosed basal cell adenoma in head and neck between Mar 2007 and Jan 2016 in our department were analyzed retrospectively. The ratio of male 3 to female 6 was 1:2. The median age of the patients was 48.9 years old(22 to 65 years). 5 cases affected parotid gland, 1 occurred in left maxillary sinus and infratemporal fossa, 1 involved nasopharyngeal and pterygopalatine fossa, 1 originated from nasal vestibule and 1 derived from nasal septum. RESULTS 8 of the patients underwent surgical treatment, while one patient with tumor involving the left maxillary sinus and infratemporal fossa was given a transnasal surgery for concurrent rhinosinusitis and subsequently confirmed by pathology. The postoperative follow-up period was between 1 and 10 years. One patient with tumor affecting infratemporal fossa recurred 1.5 years after surgery, while the rest shown no signs of recurrence and complication. CONCLUSION Basal cell adenoma in head and neck is a rare kind of disease. Clinical features and imaging helped to differenced basal cell adenoma in head and neck from other diagnoses, but definite diagnosis relies on the pathological tests.Surgery may provide good effects and prognosis on patients with basal cell adenoma.

Objective To study the analysis of auditory rehabilitation outcomes of patients with cochlear nerve canal stenosis after cochlear implantation(CI).Methods A cohort of 30 patients with bilateral profound senso-rineural hearing loss who were diagnosed with cochlear neural canal stenosis by high-resolution CT were tested with evoked compound action potential (ECAP)and evoked auditory brainstem response (EABR)during and 3 ,6 , 9 months after CI.Audiometry in sound field was also assessed before and 3 ,6 ,9 months after CI.Among the co-hort,1 7 patients over 3 years old underwent postoperative speech recognition rate test.All the auditory rehabilita-tion outcomes were analyzed.Results ① For all 30 patients,there were no obvious differences of ECAP and EABR waveforms tested in 3,6 and 9 months after CI.②The thresholds in sound field in 3,6,9 months after CI were 65 ±8 dB HL,62 ±4 dB HL and 61 ±7 dB HL,respectively.The thresholds in sound field were significantly im-proved after than before CI (100 ±5 dB HL).③ The single vowel recognition rates of 17 patients in 3 ,6 and 9 months after CI were 55%±7%,56%±8% and 80%±4%,respectively.The single vowel recognition rate was significantly improved in 9 months after than before CI(52%±8%).The single consonant recognition rates of 17 pa-tients in 3 ,6 and 9 months after CI were 9%±3%,8%±4% and 9%±2%,respectively.The single consonant recognition rates were not significantly improved after than before CI (8%±2%).Conclusion ① For patients with bi-lateral cochlear neural canal stenosis,neither ECAP nor EABR waves were produced during or after CI.The language com-munication of patients is limited as a result of their poor subjective thresholds in sound field and speech recognition rates.

Objective:To discuss the influence of intensive lipid-lowering therapy by atorvastatin on blood lipid and serum von Wille-brand factor ( vWF) and thrombomodulin ( TM) levels of patients with unstable angina pectoris. Methods: Totally 88 cases of patients with unstable angina pectoris were selected and divided into the intensive group (n=44) and the routine group (n=44) at random. The patients in the two groups were given routine medical treatment, such as nitrates, bayaspirin,β-blocker, low molecular heparin and etc. The patients in the routine group were orally given atorvastatin 20mg, qd, while the patients in the intensive group were given atorvastatin 40mg, qd with the treatment course of 8 weeks. The changes in serum vWF and TM levels in the two groups before and after the medical treatment were observed, and the occurrence rates of cardiac ischemia related events and untoward effect during the medical treatment were compared as well. Results:After the 8-week medical treatment, the levels of TC, TG and LDL-C in the two groups were obviously declined and the levels of HDL-C were obviously increased than those before the treatment (P<0. 05 or P<0. 01), and the declining and increasing rates in the intensive group were much higher than those in the routine group (P<0. 05). The serum vWF and TM levels in the two groups were obviously declined than those before the treatment (P<0. 05 or P<0. 01), and the declining rates in the intensive group were much higher than those in the routine group (P<0. 05). The occurrence rates of cardiac ischemia related events in the inten-sive group during the medical treatment were much lower than those in the routine group (P<0. 05). Respectively 4 and 6 cases of unto-ward effect appeared in the routine group and the intensive group during the medial treatment with light symptom, and the difference showed no obvious statistical significance (P>0. 05). Conclusion:Atorvastatin intensive lipid-lowering has favorable curative effect and security in the treatment of unstable angina pectoris, which can reduce the occurrence rates of cardiac ischemia related events, and the mechanism is related to reducing blood lipid and serum vWF and TM levels and improving the function of vascular endothelial cells.

Objective:To evaluate the efficacy and safety of palonosetron in preventing chemotherapy-induced vomiting. Meth-ods:A multi-center, randomized, double-blind, and self-cross-over positively controlled clinical trial design was used. All patients were randomized into two groups, as follows:Regiment A (61 cases) and Regiment B (64 cases). Regimen A with palonosetron hydrochlo-ride injection (test agent) was used in the treatment cycle A, whereas granisetron hydrochloride injection (control drug) was used in the cycle B. Treatments were randomly administered on the patients of the two groups. Regimen B was on the contrary, the control drug was used in the cycle A, and the test agent was used in the treatment cycle B. All patients treated with the test agent were classified as the test group, whereas those treated with the control drug were classified as the control group. Complete control rate and adverse reac-tion of acute and delayed vomiting in the two groups during the two cycles of chemotherapy regimen were compared. Results: In Group One, the complete control rate of delayed vomiting was significantly higher in the palonosetron administration cycles than in the granisetron cycles (76.92%vs. 55.38%, P=0.0110). In the same group, the frequency of vomiting was significantly less in palonosetron cycles than in the granisetron cycles during day 1 to day 5 (1.32±3.42 vs. 1.94±3.03, P=0.0096). The incidences of adverse effects were low in both groups. No grades 3 and 4 adverse effects were observed. Conclusion: Palonosetron showed efficacy in preventing the acute and delayed chemotherapy-induced vomiting. The drug is superior to granisetron, specifically in delaying vomiting in Group One. Palonosetron hydrochloride showed slight adverse effects. Hence, this drug can be used in clinic.

Objective To explore the effects of X-ray irradiation combined with RNAi against signal transducer and activator of transcription 3(STAT3)on the radiosensitivity of human esophageal carcinoma cells.Methods Human esophageal carcinoma cells of the line Eca-109 were euhured.Three pairs of DNA template aiming at the base sequences of the coding regions 2037-2055,1243-1261,and 455-473 of the STAT3 mRNA were synthesized(siRNAI,siRNA2,and siRNA3),and a negative sequence was synthesized to be used as control.STAT3-siRNA positive recombinant plasmids(pRNAT-U6.1-siRNAI,pRNAT-U6.1-siRNA2, and pRNAT-U6.1-siRNA3), and a STAT3-siRNA negative recombinant plasmid (pRNAT-U6.1-negative)were thus constructed and then transfected into the cultured Eca-109 cells,which were divided into transfection reagent control group,pRNAT-U6.1-siRNAl-3 transfection groups,and pRNAT-U6.1-negative centrel group.The positive eell clones were screened.RT-PCR and Westem blotting were used to detect the STAT3 mRNA and protein expression.The transfected Eca-109 cells were exposed to 0,2,4,6,and 8 Gy of X-rays,respectively,and the survival fraction of the cells was analyzed by clone formation assay.Flow cytometry was applied to analyze the cycle arrest and cell apoptosis 4 Gy post-irradiation.Results Agarose gel electrophoresis confirmed the successful construction of the plasmid pRNAT-U6.1-siRNA.RT-PCR and Western blotting demonstrated that the mRNA and protein expression levels of STAT3 transfected with sTAT3-siRNA3 were both significanfly lower than those of the control groups.At 2-8 Gy, the survival fractions of the siRNA3 group were aU significantly lowered than those of the control group(t＝-0.228--0.051,P<0.05).Flow cytometry showed that the percentage of the cell cycle G0/G1 phase and the apoptosis rate of the siRNA3 group were both significantly higher than those of the control groups at 4 Gy post-irradiation(t＝-13.137-16.350,P<0.01).Conclusions X-ray irradiation combined with RNAi against sTAT3 could inhibit the proliferation of the human esophageal carcinoma cells,induce cell cycle arrest and apoptosis,improve the radiosensitivity in Eta-109 cells.

BACKGROUND: At present, studies on repair of cartilage defect have been focused on tissue engineering technique. Growth factors are one of the most important parts. However, the effect and security of growth factors have not been confirmed. Studies have shown that Weilingxian can maintain and promote the synthesis of proteoglycan, collagen Ⅱof chondrocyte, and it also can promote proliferation of chondrocyte and expression of transforming growth factor (TGF)-β1 mRNA.OBJEGTIVE: To investigate the feasibility of injectable chitosan/β-glycerol phosphate (C/β-GP) encapsulating allograft chondrocytes on the repair of articular cartilage defects and the intervention effect and possible mechanisms of Weilingxian.METHODS: A 0.4-mm defect was established on knee articular cartilage. Expeirmental New Zealand rabbits were randomly divided into three groups: Weilingxian, common culture media, and model groups. In the common culture media group, the samples were treated with C/β-GP and chondrocyte suspension (1 mL); at 2 days after gel injection, Weilingxian or common culture media (1 mL) were respectively given into joint cavity, once a day, for 7 successive days. The samples in the model group were not treated. Gross, histological (HE staining, TB staining), type Ⅱ collagen immunohistochemical, and Wakitani score examinations were performed on 6 and 12 weeks after surgery.RESULTS AND CONCLUSION: Defects of articular surface were well filled in Weilingxian and common culture media groups, and hyaline cartilage-like structure was formed. The surface flatness and degree of integration with surrounding tissue of Weilingxian group was better than common culture media group. Formation of cartilage-like and secretion of cartilage matrix and specificity of collagen type Ⅱ were found in histological slices. Defects in the model group were not repaired, while tissue proliferative degeneration was observed. Integration of.repair tissue with surrounding tissue, histology and amount of type Ⅱ collagen secretion in Weilingxian group were better than common culture media group. Wakitani scores of Weilingxian group and common culture media group were significantly lower than model group (P < 0.01), andscores of Weilingxian group was significantly lower than common culture media group (P<0.05). Injectable chitosan/β-glycerolphosphate gel encapsulating allograft chondrocytes could repair articular cartilage defects, and Weilingxian was able to promote the process of it, this manifested the role like growth factor in tissue engineering technique repairing articular cartilage defects.

BACKGROUND:Studies have shown that Weilingxian can maintain and promote the synthesis of proteoglycan and collagen Ⅱ of chondrocyte,and protect artlcular cartilage and postpone the development of osteoarthdtis by inhibiting the level of intedeukin-1(1L-1)possibly.OBJECTIVE:Based on the previous studies,to observe the effect of Weilingxian on the proliferation of rabbit knee articular chondrocyte and transforming growth factor-β1 mRNA expression,and then to explore the role and possible mechanism of Weilingxian in the treatment of osteoarthdtis.METHODS:Knee cartilage was shredded after harvested from New Zealand white rabbits under sterile conditions,and chondrocytes were isolated and cultured by the way Of enzymatic digestion.After identifying by toluidine blue staining,the third-passage calls in the logarithmic growth phase were cultured in vitro and randomly divided into two groups after adherence.The experimental groups were cultured in DMEM with 0.01,0.05,0.1,0.5,and 1.0 mg/mL Weilingxian,while the control group was given with normal medium alone.Chondrecytes morphology was observed under an inverted phase contrast microscope,and the phenotype was identified by toluidine blue staining;Methyl Thiazolyl Tetrazolium(MTT)assay method was adopted to observe the influenca of Weilingxian with difierent concentrations on the proliferation of chondrocytes,and anti-transcription-polymerase chain-type reaction(RT-PCR)was used to assay the expression changes of transforming growth factor-β1 mRNA.RESULTS AND CONCLUSlON:Primary cultured chondrocyte was round-shaped,and most of It adhered after 24 hours,the appearance was polygonal and irregular-shaped;after passage,cell growth was faster than before,the typical appearance was slabstone-like;long spindle-shaped chondrocytes appeared after four generations;after six generations,most cells showed long spindle-shaped fibroblast-like appearance,the rate of growth also slowed down.Extracellular matrix of chondrocytes was stained to be blue by toluidine blue staining,and the nucleus was dark blue.Different concentrations of Weilingxian could promote the proliferation of chondrocytes,effect of 0.5 mg/mL group was significantly,and the peak of proliferation was on the third day.0.05,0.1,0.5,and 1.0 mg/mL Weilingxian group could promote the expression of transforming growth factor-β1 mRNA.and there was no significant difference between four groups(P＞0.05),but the peak was at 0.5 mg/mL group.Weilingxian can promote proliferation of chondrocyte and transfonlling growth factor-β1 mRNA expression,and these may be one of the possible mechanisms that Weilingxian can work in the treatment of osteoarthritis.