Objective:To investigate the possible role and mechanism of purinergic ligand-gated ion channel 7(P2X7)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3) inflammasome pathway in cognitive impairment induced by sleep deprivation (SD)mice.Methods:SPF grade male C57BL / 6J mice aged 6-8 weeks were randomly divided into 3 groups according to the random number table method with 6 mice in each group.They were normal control group (CC group), SD group and SD+ P2X7 receptor antagonist brilliant blue G(BBG) group (SD+ BBG group). Modified multiple platform method was used to establish a 5-day SD model in mice.During the SD intervention period, the mice in SD+ BBG group were injected with BBG(50 mg/kg) intraperitoneally once a day, while the mice in CC group and SD group were injected with the same volume of 0.9% sodium chloride solution.Morris water maze was conducted to evaluate the cognitive function of mice.The protein expression levels of P2X7, NLRP3, caspase-1, apoptosis-associated proteins(ASC) and interleukin-1β(IL-1β) in hippocampus were detected by Western blot.RT-qPCR was used to detect the mRNA expression levels of tumor necrosis factor-α(TNF-α), IL-1β, interleukin-18(IL-18) and microglial polarization surface markers CD206 and CD86 in hippocampus.Graph pad Prism 8.0 software and SPSS 25.0 software were used for statistical analysis and mapping.Results:(1) The interaction effect between time and groups of escape latency in three groups of mice was significant ( F=15.76, P<0.001). From the 2nd to 5th day, the escape latencies of mice in SD group were higher than those of CC group, while the escape latencies of mice in SD+ BBG group were lower than those of SD group (all P<0.05). (2)The results of the space exploration experiment showed that there were statistically significant differences in target quadrant residence time and the times of crossing the platform( F=6.65, P=0.009; F=12.39, P<0.001). The target quadrant residence time ((23.42±0.55) s) and times of crossing the platform ((17.67±0.71) times) of the SD group were both lower than those of the CC group ((29.48±1.78) s, (23.33±0.95) times) (both P<0.05), while the target quadrant residence time ((28.62±1.19) s) and the times of crossing the platforms ((21.33±0.76) times) of the SD+ BBG group were both higher than those of the SD group (both P<0.05). (3)There were statistically significant differences in the protein levels of inflammatory related proteins such as P2X7, NLRP3, caspase-1, ASC and IL-1β in the hippocampus of mice among the 3 groups( F=8.23, 8.97, 8.45, 54.42, 8.12, all P<0.05). Compared with CC group, the protein levels of P2X7 ((0.93±0.02), (0.71±0.04)), NLRP3 ((0.97±0.04), (0.62±0.09)), caspase-1 ((1.00±0.03), (0.76±0.07)), ASC ((0.96±0.02), (0.77±0.04)) and IL-1β ((0.85±0.07), (0.54±0.04)) in SD group were all higher (all P<0.05). Compared with SD group, the protein levels of P2X7 (0.74±0.05), NLRP3 (0.78±0.02), caspase-1 (0.74±0.04), ASC (0.67±0.02), IL-1β (0.53±0.07) in SD+ BBG group were all lower (all P<0.05). (4)There were statistically significant differences in the mRNA levels of IL-18, IL-1β, TNF-α, CD86 and CD206 in hippocampus among the three groups ( F=12.80, 12.28, 105.80, 7.06, 30.19, all P<0.05). The mRNA levels of IL-18, IL-1β, TNF-α, CD86 in SD group were all higher than those in CC group(all P<0.05), while the mRNA level of CD206 in SD group was lower than that in CC group( P<0.05). Compared with SD group, the mRNA levels of IL-18, IL-1β, TNF-α, CD86 were lower in SD+ BBG group (all P<0.05), while the CD206 mRNA level of SD+ BBG group was higher than that in SD group( P<0.05). Conclusion:SD intervention can lead to cognitive impairment and increased expression of P2X7 in hippocampus of mice, which may be related to the activation of P2X7/ NLRP3 inflammasome signaling pathway, promoting the polarization of microglia into pro-inflammatory type and up-regulating the expression of pro-inflammatory cytokines.Inhibition of P2X7 can improve the cognitive function of mice.

Objective:To investigate the characteristics of transcranial sonography (TCS) in patients with restless legs syndrome (RLS), and analyze the correlations of scores of RLS Self-rating Severity Scale by International Restless Leg Syndrome Study Group (IRLS) and TCS parameters with clinical data of these patients.Methods:Twenty-one patients with RLS admitted to the Sleep Disorder Clinic of our hospital from September 2020 to January 2021 were selected as RLS group, and 23 healthy controls at the same time period were recruited as control group. IRLS was used to evaluate the severity of patients in the RLS group, and the 14-item Hamilton anxiety rating scale (HAMA-14) and 24-item Hamilton depression rating scale (HAMD-24) were used to evaluate the anxiety and depression of subjects from the 2 groups. Pittsburgh Sleep Quality Scale (PSQI), Insomnia Severity Index (ISI) and Epworth Sleepiness Scale (ESS) were used to evaluate the sleep quality of subjects from the 2 groups. TCS was used to examine the occurrence of hypoechoic substantia nigra and raphe nucleus rupture and the width of the third ventricle in the two groups. The clinical data and TCS parameters of patients in the 2 groups were compared, and the correlations of IRLS scores and TCS parameters with clinical features of patients in the RLS group were analyzed.Results:As compared with those in the control group, the HAMA-14, HAMD-24, ISI and PSQI scores in the RLS group were statistically higher ( P<0.05). As compared with the control group, RLS group had significantly higher proportion of patients with hypoechoic substantia nigra or raphe nucleus rupture ( P<0.05). In RLS patients, the IRLS scores were positively correlated with HAMA-14, HAMD-24, and ISI scores ( P<0.05); ESS scores were negatively correlated with hypoechoic substantia nigra and width of the third ventricle ( rs=-2.005, P=0.045; r=-0.477, P=0.029); width of the third ventricle was negatively correlated with gender (male) and years of education ( rs=-0.592, P=0.005; r=-0.627, P=0.002), and positively correlated with age and course of the disease ( r=0.756, P<0.001; r=0.167, P=0.047). Conclusions:Patients with RLS are prone to anxiety, depression and sleep disorders; their TCS shows hypoechoic substantia nigra and raphe nucleus rupture. RLS severity may affect HAMA-14, HAMD-24, and ISI scores. Gender, age, years of education, course of disease, and ESS scores of RLS patients may affect TCS related parameters.

Objective:To investigate the correlation between single nucleic acid polymorphisms (SNPs) of MEIS1, BTBD9, MAP2K5, PTPRD and restless leg syndrome (RLS).Methods:By searching the literatures published before March 1, 2021 at home and abroad, case-control studies on risk genes associated with RLS were collected, and the Review Manager 5.3 and Stata 15.1 softwares were used for statistical analysis.Results:A total of 8 studies were included, with a total of 7 824 cases and 14 645 controls.Meta analysis results showed that the SNPs locus of the risk gene associated with RLS was MEIS1 rs2300478( OR=1.68, 95% CI: 1.59-1.78), BTBD9 rs9296249( OR=1.62, 95% CI: 1.47-1.77), BTBD9 rs9357271( OR=1.49, 95% CI: 1.44-1.55), MAP2K5 rs12593813( OR=1.44, 95% CI: 1.36-1.53), MAP2K5 rs11635424( OR=1.47, 95% CI: 1.34-1.60)and PTPRD rs1975197( OR=1.34, 95% CI: 1.21-1.49). Conclusion:MEIS1 rs2300478, BTBD9 rs9296249, BTBD9 rs9357271, MAP2K5 rs12593813, MAP2K5 rs11635424 and PTPRD rs1975197 are the risk loci of RLS.