Objective To investigate the role of transcription factor EB(TFEB)in endotoxin-tolerant macrophages.Methods The RAW264.7 cells were divided into blank group(DMEM medium),LPS 5 group(5 ng/mL LPS treatment for 4 h),LPS 100 group(100 ng/mL LPS treatment for 4 h),and tolerance group(5 ng/mL LPS for 12 h followed by 100 ng/mL LPS for 4 h).The releases of inflammatory factors TNF-α and IL-6 were measured using ELISA.Western blotting and immunofluorescence assay were used to evaluate the distribution of autophagy-related proteins LC3 and P62,as well as TFEB in the cytoplasm and nucleus.Lentiviral overexpression of TFEB or siRNA-mediated knockdown of TFEB were performed to observe the changes in autophagy levels and bacterial clearance ability in the tolerant cells.Results The cells in the tolerance group had significantly lower contents of TNF-α and IL-6,as well as reduced bacterial clearance ability(P<0.01),down-regulated LC3 expression while up-regulated P62 level,and decreased expression of TFEB in both the cytoplasm and nucleus(P<0.01)when compared with the cells of the LPS 100 group.Overexpression of TFEB significantly increased LC3 level,reduced P62 level,and enhanced bacterial clearance ability in the endotoxin-tolerant cells(P<0.01).In contrast,siRNA-mediated knockdown of TFEB had no significant impacts on LC3 and P62 expression levels or bacterial clearance ability.Conclusion Overexpression of TFEB can restore the autophagy of endotoxin-tolerant cells and enhance their bacterial clearance capacity,thereby alleviating the immunosuppressive state of sepsis.These findings suggest that TFEB holds promise as a potential therapeutic target for the prevention and treatment of sepsis.

OBJECTIVE To establish the HPLC fingerprint and content determination of five components in Ranunculus sceleratus L.. METHODS The separation was developed on an Agilent ZORBAX SB C18 chromatographic (4.6 mm×250 mm, 5 μm)column by gradient elution with methanol(A)-0.1 % phosphoric acid aqueous solution(B) as mobile phase to establish HPLC fingerprint of Ranunculus sceleratus L.. Combined with similarity evaluation, cluster analysis, principal component analysis, and orthogonal partial least squares-discriminant analysis, the quality of 13 batches of Ranunculus sceleratus L. was evaluated. RESULTS Thirteen batches of Ranunculus sceleratus L. samples were calibrated with 20 common peaks, of which 5 common peaks were identified, and the similarity ranged from 0.874 to 0.984. The results of cluster analysis and principal component analysis were basically the same, indicating that there might be differences in the content of chemical components of Ranunculus sceleratus L. in different regions. Protocatechuic aldehyde, caffeic acid, ferulic acid, hyperoside and isoquercitrin were determined in thirteen batches of Ranunculus sceleratus L., and their contents were 0.016−0.035, 0.010−0.070, 0.010−0.029, 0.016−0.051, 0.028−0.086 mg·g–1, respectively. CONCLUSION The established HPLC fingerprint and content determination method is simple, stable, and reliable, which can be used for qualitative analysis and provide reference to quality evaluation and resource utilization of Ranunculus sceleratus L..

Objective To investigate the popularization of cardiopulmonary resuscitation(CPR)knowledge and science popularization needs among urban and rural residents in Tonghai County,Yuxi City,Yunnan Province,so as to explore the establishment of an efficient and appropriate science popularization model.Methods A total of 300 residents aged 15-60 years old were selected from Tonghai County,Yuxi City,Yunnan Province using stratified and simple random sampling methods.A self-designed questionnaire was used to conduct an anonymous questionnaire survey.Results Only 20.3%of Tonghai County residents master CPR skills,and 26.2%of Tonghai County residents have never heard of CPR.There is a statistically significant difference in the awareness rate of CPR between rural residents and non-rural residents(P<0.01).There are differences in residents'age and CPR awareness(P<0.01),the age and CPR are inversely proportional.The residents have a higher willingness to perform chest compressions and mouth-to-mouth resuscitation on strangers,66.2%and 68.6%respectively.63.79%of residents have never attended relevant training.But 92.76%of the people said they were willing to participate in the relevant training when they learned the training news.Conclusion Residents in Tonghai County generally lack knowledge of CPR first aid,but the demand for first aid knowledge of residential CPR is high and the attitude towards rescue is positive.It is recommended that relevant departments increase CPR science popularization and training efforts,and popularize CPR into villages.

Objective:To study the effects of the combined application of NaF and Nisin on Streptococcus mutans(S.mutans).Meth-ods:The minimum inhibitory concentration(MIC)of Nisin and NaF against S.mutans was determined by microdilution method respec-tively.The fractional inhibitory concentration(FIC)was calculated by checkerboard method.The inhibition effect of the combination of NaF and Nisin at(NF)the same MIC concentration on acid production and acid resistance of S.mutans was detected.Crystal violet staining was used to detect the effects of NF in the inhibition of the biofilm formation,and the damage and dispersion of the established biofilms.The changes of the biofilms were observed by CLSM.Results:The MIC of NaF and Nisin was 0.6 mg/mL and 20 mg/mL re-spectively.The FIC was 0.75.The 1/8×MIC NF showed significantly higher inhibition on acid production and biofilm formation than 1/4×MIC NaF or Nisin(P＜0.05),but it has no obvious dispersion effect on established biofilms.The 1/2×MIC NF showed stronger in-hibition effect on the acid resistance of S.mutans in the membrane than 1×MIC NaF or Nisin(P＜0.05).At the concentration of 2x MIC,any component didn't cause obvious damage on the established biofilm structure.Conclusion:Nisin and NaF have synergistic in-hibitory effects on the proliferation,acid production,acid resistance and biofilm formation of S.mutans.

Objective:To explore the association between cognitive function and the level of gamma-amino-butyric acid(GABA)in anterior cingulate cortex(ACC)before and after treatment in patients with major depres-sion disorder.Methods:Totally 31 medication-naive patients with major depression disorder meeting the criteria of the Diagnostic and Statistical Manual of Mental Disorders,Fifth Edition(DSM-5)and 33 normal controls were col-lected.Each eligible patient received treatment with selective serotonin reuptake inhibitor agents for 8 weeks.The MATRICS Consensus Cognitive Battery(MCCB)was used to evaluate the cognitive function.By means of 1H magnetic resonance spectroscopy,anterior cingulate cortex GABA concentrations were measured.Results:At base-line,the concentration of ACC GABA relative to water(GABA+/W)was lower in the patient group than in the control group(P＜0.05)and increased after treatment(P＜0.05).ACC GABA+/W was negatively associated with verbal learning and visual memory score in patient group at baseline(correlation coefficient and P value were r=-0.40,P＜0.05;r=-0.42,P＜0.05,respectively).The ACC GABA+/W difference resulted of treatment in patient group was positively associated with the difference of working memory score and the difference of reasoning and problem-solving score(correlation coefficient and P value were r=0.58,P＜0.05;r=0.66,P＜0.05,respec-tively).Conclusion:The cognitive dysfunction of patients with major depression disorder may not be related to the degree of depression and anxiety.And improvement of cognitive function may be associated with increase of ACC GABA concentrations.

Objective:To investigate the relationship between niacin skin reaction sensitivity and cognitive function by comparing the difference of niacin skin flushing response and cognitive function between the patients with depression and the normal control group.Method:A cross-sectional study was conducted to recruit 40 patients with depression who met the diagnostic criteria of DSM-Ⅳ (American Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition) at The Second People′s Hospital of Zhumadian from July 2018 to August 2019. In the meanwhile, 32 gender-and-education-matched patients were recruited and allocated into the control group. General demographic and clinical data were collected. Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) was used to test individual′s cognitive functions. A Swedish laser Doppler blood flow meter, PeriFlux 5000, was used to test the skin flushing reaction of forearm niacin at eight concentrations (10 -5 mol/L, 10 -4 mol/L, 10 -3.5 mol/L, 10 -3 mol/L, 10 -2.5 mol/L, 10 -2 mol/L, 10 -1.5 mol/L, and 10 -1 mol/L) gradient. The difference of blood flow values in 8 concentration gradients between the two groups was analyzed by repeated measurement multiple comparison and simple effect. The MBF value and LogEC50 were analyzed by non-parametric test. Spearman rank correlation analysis was used to analyze the correlation between LogEC50 and cognitive function scores. Result:In the skin flushing reaction of niacin test, when the concentration of niacin solution was 10 -3.5 mol/L and 10 -3mol/L, there was a significant difference in Blood Flow (BF) between patients with depression and normal subjects ( t=-0.490,-4.626, P<0.01, respectively). The BF value of the normal control group was higher than that of depressed patients. LogEC50 was significantly different between the two groups ( t=4.196, P<0.01). LogEC50 of the normal control group (-3.513±0.848) was lower than that of the depression group (-2.776±0.568). The difference in RBANS between the two groups was tested by independent sample t test. Compared with the normal control group, the scores of the depressed group were significantly lower on the test of list learning ( t=-2.127, P=0.037), coding test ( t=-3.226, P=0, 002), list recall ( t=-3.697, P<0.01), story recall ( t=-2.687, P=0.009), and figure recall ( t=-2.660, P=0.010). There was a negative correlation between list recall ( r=-0.407, P=0.032), story recall ( r=-0.376, P=0.049) and figure recall ( r=-0.455, P=0.015) in the depression group. Maximal blood flow (MBF) was positively correlated with figure copying ( r=0.428, P=0.023). Conclusion:The niacin sensitivity and the score of cognitive function of depression patients are lower than those of the normal control group, and the decrease of cognitive function of depression patients may be related to the disorder of phospholipase A2 (PLA2) signaling pathway.

Objective:To investigate the relationship between niacin skin reaction sensitivity and cognitive function by comparing the difference of niacin skin flushing response and cognitive function between the patients with depression and the normal control group.Method:A cross-sectional study was conducted to recruit 40 patients with depression who met the diagnostic criteria of DSM-Ⅳ (American Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition) at The Second People′s Hospital of Zhumadian from July 2018 to August 2019. In the meanwhile, 32 gender-and-education-matched patients were recruited and allocated into the control group. General demographic and clinical data were collected. Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) was used to test individual′s cognitive functions. A Swedish laser Doppler blood flow meter, PeriFlux 5000, was used to test the skin flushing reaction of forearm niacin at eight concentrations (10 -5 mol/L, 10 -4 mol/L, 10 -3.5 mol/L, 10 -3 mol/L, 10 -2.5 mol/L, 10 -2 mol/L, 10 -1.5 mol/L, and 10 -1 mol/L) gradient. The difference of blood flow values in 8 concentration gradients between the two groups was analyzed by repeated measurement multiple comparison and simple effect. The MBF value and LogEC50 were analyzed by non-parametric test. Spearman rank correlation analysis was used to analyze the correlation between LogEC50 and cognitive function scores. Result:In the skin flushing reaction of niacin test, when the concentration of niacin solution was 10 -3.5 mol/L and 10 -3mol/L, there was a significant difference in Blood Flow (BF) between patients with depression and normal subjects ( t=-0.490,-4.626, P<0.01, respectively). The BF value of the normal control group was higher than that of depressed patients. LogEC50 was significantly different between the two groups ( t=4.196, P<0.01). LogEC50 of the normal control group (-3.513±0.848) was lower than that of the depression group (-2.776±0.568). The difference in RBANS between the two groups was tested by independent sample t test. Compared with the normal control group, the scores of the depressed group were significantly lower on the test of list learning ( t=-2.127, P=0.037), coding test ( t=-3.226, P=0, 002), list recall ( t=-3.697, P<0.01), story recall ( t=-2.687, P=0.009), and figure recall ( t=-2.660, P=0.010). There was a negative correlation between list recall ( r=-0.407, P=0.032), story recall ( r=-0.376, P=0.049) and figure recall ( r=-0.455, P=0.015) in the depression group. Maximal blood flow (MBF) was positively correlated with figure copying ( r=0.428, P=0.023). Conclusion:The niacin sensitivity and the score of cognitive function of depression patients are lower than those of the normal control group, and the decrease of cognitive function of depression patients may be related to the disorder of phospholipase A2 (PLA2) signaling pathway.

Objective:To estimate the infection rate of close contacts of COVID-19 cases, and to evaluate the risk of COVID-19 under different exposure conditions.Methods:A prospective study was used to conduct continuous quarantine medical observations of close contacts of people infected with COVID-19, collect epidemiological, clinical manifestations, and laboratory test data to estimate the infection rate of close contacts under different exposures.Results:The epidemiological curve of COVID-19 in Ningbo showed persistent human-to-human characteristics. A total of 2 147 close contacts were tracked and investigated. The total infection rate was 6.15%. The infection rates of confirmed cases and positive contacts were 6.30% and 4.11%, respectively. The difference was not statistically significant ( P>0.05). Among close contacts of different relationships, friends/pilgrims (22.31%), family members (18.01%), and relatives (4.73%) have a higher infection rate, and close contacts of medical staff were not infected. Differences in infection rates among the close contacts were statistically significant ( P<0.005). Living with the case (13.26%), taking the same transportation (11.91%), and dining together (7.18%) are high risk factors for infection. Cross-infection in the hospital should not be ignored (1.94%). The median of incubation period is 5 days. Conclusion:The infection rate of close contacts of COVID-19 cases is high, and isolation medical observation measures should be implemented in strict accordance with the close contact management plan.

Objective:To estimate the infection rate of close contacts of COVID-19 cases, and to evaluate the risk of COVID-19 under different exposure conditions.Methods:A prospective study was used to conduct continuous quarantine medical observations of close contacts of people infected with COVID-19, collect epidemiological, clinical manifestations, and laboratory test data to estimate the infection rate of close contacts under different exposures.Results:The epidemiological curve of COVID-19 in Ningbo showed persistent human-to-human characteristics. A total of 2 147 close contacts were tracked and investigated. The total infection rate was 6.15%. The infection rates of confirmed cases and positive contacts were 6.30% and 4.11%, respectively. The difference was not statistically significant ( P>0.05). Among close contacts of different relationships, friends/pilgrims (22.31%), family members (18.01%), and relatives (4.73%) have a higher infection rate, and close contacts of medical staff were not infected. Differences in infection rates among the close contacts were statistically significant ( P<0.005). Living with the case (13.26%), taking the same transportation (11.91%), and dining together (7.18%) are high risk factors for infection. Cross-infection in the hospital should not be ignored (1.94%). The median of incubation period is 5 days. Conclusion:The infection rate of close contacts of COVID-19 cases is high, and isolation medical observation measures should be implemented in strict accordance with the close contact management plan.

Objective To investigate the functional connection(FC) between the anterior cingulate cortex (ACC) and other brain regions in unipolar depression(UD) and bipolar depression(BD), and the correlation to the severity of depressive symptoms. Methods 30 patients who met the diagnosis criteria of unipolar depression in DSM?Ⅳ and another 30 patients who met the diagnosis criteria of bipolar depression in DSM?Ⅳ were matched with 32 healthy controls(HC) by gender?, age?, and education. All subjects completed the resting?state functional magnetic resonance scans. After the imaging preprocessing, seed?based functional connectivity analysis was performed on the bilateral ACC. Pearson correlation analysis was performed between the FC values and the severity of depressive symptoms (measured by HAMD17 scores). Results (1) Compared to the health controls, the connectivity between the left ACC (LACC) and the left triangular part of the inferior frontal gyrus(t=3.48) and the connectivity from the LACC to the left inferior parietal gyrus(t=3.15) in patients with unipolar depression significantly increased, so did the connectivity between the right ACC(RACC) and the left triangular part of the inferior frontal gyrus(t=3.52); (2) Compared to the healthy controls, the connectivity from LACC to the right middle temporal gyrus (t=-4.00) in patients with bipolar depression significantly decreased, so did the connectivity between RACC and the other regions induding the left postcentral(t=-3.46), the right superior temporal gyrus(t=-2.86) and the right middle temporal gyrus(t=-3.40);(3)Compared to the patients with bipolar depression,patients with unipolar depression increased connectivity between the LACC and the other regions including the left triangular part of the inferior frontal gyrus(t=3.58), the right triangular part of the inferior frontal gyrus(t=4.06), the right middle temporal gyrus (t=3.25), the left inferior parietal gyrus(t=3.27), and the right inferior parietal gyrus(t=3.99). While the RACC increased its connectivity to the left triangular part of the inferior frontal gyrus(t=3.13), the left inferior parietal gyrus(t=3.42), and the right inferior parietal gyrus(t=3.78) (Alphasim?corr, P<0.001); (4) The severity of depressive symptoms were found uncorrelated to the FC value changes among the different regions(-0.20.05). Conclusion The results suggest that there are much difference in the brain function activity between UD and BD. The connectivity between ACC and other regions is stronger in UD patients, while weaker in BD patients, therefore the FC value could be possibly explored as an independent future biomarker to distinguish BD from UD regardless the symptoms severity.