Objective To observe the effect of the nuclear factor (NF)-κB inhibitor on the inflammatory injury and the secondary remote damage in remote areas of the CA1 region in the right hippocampus of the focal cerebral ischemic/reperfusion rats,and the NF-κB essential modifier binding domain (NBD) peptide was used to inhibit the activation of NF-κB signaling pathway to explore the function and mechanism of the NBD peptide in restraining inflammatory injury and reducing secondary remote damage in the hippocampus CA1 region.Methods According to the random number table,the male Sprague-Dawley (SD) rats were randomly divided into a sham group (n =24),an ischemia/reperfusion (I/R)group (n =38),a NBD group (n =38) and a modified type peptide (MT-NBD) group (n =38),then at the time point of 24 h and 7 d after reperfusion,the above 4 groups were divided into 2 subgroups.The experimental models were made by middle cerebral artery occlusion (modified line plug method) for 2 hours.The NBD peptide and the MT-NBD peptide were respectively injected into the right hippocampus of the experimental groups.The injury of neurons was examined by the methods of H&E and Fluoro-Jade B-(FJB)staining.The levels of interleukin-1β (IL-1β) and IL-1Ra were detected by using enzyme-linked immunosorbent assay.The protein expressions of NF-κB p65 and IκBα were analyzed by Western blotting and the double-labelling immunofluorescence.Results Compared with the NBD group (24 h 0.206 ±0.013,7 d 0.090 ±0.012) and the sham group (24 h 0.120 ±0.007,7 d 0.100 ±0.014),the NF-κB p65 protein expression in the I/R group (24 h 2.340 ± 0.101,7 d 2.440 ± 0.081) was increased significantly (q =64.431,66.704,67.747,56.624,all P ＜ 0.05).The level of IL-1β was remarkably increased in the I/R group (24 h (1.850 ±0.192) ng/ml,7 d (1.000 ±0.178) ng/ml) compared with the NBD group (24 h (1.250 ± 0.211) ng/ml,7 d (0.560 ± 0.183) ng/ml,q =10.730,9.710,P ＜0.05).The percent of survival neurons was significantly lower in the I/R group (24 h 27.50％ ± 3.59％,7 d 28.10％ ±4.46％) and the MT-NBD group (24 h 27.30％ ±4.53％,7 d 26.30％ ±5.03％)than the NBD group (24 h 58.90％ ± 3.46％,7 d 68.40％ ±4.20％,q =19.949,19.731,2.139,22.249,all P ＜0.05).The FJB staining showed that the number of neuron degeneration in the I/R group (24 h 28.10 ±2.13,7 d 29.50 ±2.45) was higher than the NBD group (24 h 12.50 ±2.41,7 d 9.30 ±2.52,q =3.211,4.521,P ＜ 0.05).Compared with the other three groups (sham group:24 h 0.130 ± 0.008,7 d 0.150 ±0.010;I/R group:24 h 1.340 ±0.213,7 d 1.750 ±0.119;MT-NBD group:24 h 1.250 ±0.114,7 d 1.620 ±0.097),the IκBα protein expression in the NBD group (24 h 1.680 ±0.148,7 d 2.010 ±0.085) was significantly increased (q =6.348,9.139,9.414,1.711,5.277,5.555,all P ＜0.05).Compared with the I/R group (24 h (0.570 ± 0.028) ng/ml,7 d (0.430 ± 0.039) ng/ml) and the MT-NBD group (24 h (0.490 ± 0.042) ng/ml,7 d (0.380 ± 0.018) ng/ml),the level of IL-1Ra in the NBD group (24 h (1.390 ± 0.055) ng/ml,7 d (1.250 ± 0.043) ng/ml) was remarkably increased (q =4.577,6.205,9.683,6.389,all P ＜ 0.05).The results between the I/R group and the MT-NBD group were not significantly different.Conclusions The research shows that NBD peptide treatment contributes to altering the NF-κB p65/IκBα expression in nucleus effectively.And it directly regulates the NF-κB activation to alleviate the inflammatory injury in the hippocampus CA1 region after the secondary remote damage.

BACKGROUND: Previous studies have confirmed that embryonic stem cells call be induced to differentiate into insulin-producing cells, but the induction process takes a long time. Most of the processes take about one month.OBJECTIVE: Activin A, all-trans retinoic acid (ATRA), basic fibroblast growth factor (bFGF) and nicotinamide were applied in vitro in combination to observe whether mouse embryonic stem cells could be induct to differentiate into insulin-producing cell in a relatively short time.DESIGN: Cell observation experiment.SETTING: Shanghai Institute of Endocrine and Metabolic Diseases, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.MATERIALS: This study was performed at Shanghai Institute of Endocrine and Metabolic Diseases from October 2004 to February 2006. Two mice of clean grade and of 12.5-14.5 days of gestational age were provided by Shanghai SLAC Laboratory Animal Co., Ltd (Permission No. 2004A034). The protocol was performed in accordance with ethical guidelines for the use and care of animals. Mouse embryonic stem cell lines were supplied by Dr Changxian Zhang (CNRS UMR5641, France). Activin A was the product of the R & D Corporation. ATRA and nicotinamide were supplied by the Sigma Corporation, USA. BFGF was supplied by Gibco Corporaion. METHODS: Head and viscera were removed from embryos of the pregnant mouse. The remaining tissues were cut into pieces and digested with trypsin. Cell suspension was centrifuged and inoculated at 3×108L-1. The cells could be used as mouse feeder layer after 2-3 times of passage. The mouse embryonic stem cells (ESCs) were inoculated onto the feeder layer in knockout Dulbecco's modified Eagle medium (DMEM) supplemented with leukemia inhibitory factors (LIF). ESCs were passaged at 1:3-1:6 after 2-3 days of culture. Culture medium with serum was added into the culture dishes to terminate the digestion. Cell fluid was centrifuged and supernatant was discarded. The sediments were prepared into suspension and inoculated at 2.5×104 with LIF-free culture medium. After 24-48 hours, embryonic bodies (EBs) were collected and replated in 1% Matrigel-coated dishes. When began to adhere to the dishes, EBs were cultured in 10% FBS/DMEM supplemented with 100μg/L activin A for 24 hours. Then EBs were switched to 10% FBS/DMEM for 6-8 hours as an interval. After this interval. EBs differentiated were cultured in 10% FBS/DMEM with 10<-6mol/L RA for another 24 hours followed by culture in 10% FBS/DMEM supplemented with 10μg/L bFGFs for 3-5 days. Finally, EBs differentiated were cultured in DMEM/F12 supplemented with N2 supplement, B27 supplement, 1μg/L laminin, 10μg/L bFGFs, and 10mmol/L nicotinamide for 3-5 days. Dithizone (DTZ) staining, inununofluorescent staining and reverse transcription-polymerase chain reaction (RT-PCR) were applied to detect insulin expression in the differentiated cells.MAIN OUTCOME MEASURES: Induction of ESCs, DTZ staining and immunofluorescent staining as wel as RT-PCR detection.RESULTS: Mouse ESCs growing on a feeder layer formed many colonies with clear boundary and dense structure. However, there was no obvious outer limit between these ESCs. EBs began to adhere to the dishes, which were coated with matrigel, on the 2nd day. After activin A and ATRA interval induction, EBs spread, and most of the living cells were epithelial cell-like when cultured in 10% FBS/DMEM supplemented with 10μg/L bFGFs. After culturing in DMEM/F12 supplemented with N2, B27, nicotinamide, bFGFs and laminin, the cells formed small clusters. The insulin-producing cells were stained dark red with DTZ, and the cells stained with primary antibody to insulin were insulin-positive. After 2 weeks of induction of activin A, ATRA, bFGFs and nicotinamide, the insulin-producing cells expressed insulin 2, Pdxl, Nkx6.1, Nkx2.2, PP, IAPP, Glut2, Somastatin, Hnf3β and Neuro D mRNA but did not express insulin 1 mRNA.CONCLUSION: Mouse ESCs call be induced to differentiate into insulin-producing cells by activin A, ATRA, bFGFs and nicotinamide in vitro. Induction time call be shortened to 2 weeks.

Objective To evaluate the effects of Cypher~(TM) Stent on endothelium regeneration and platelet activation after angioplasty. Methods Six mongrel dogs were implanted Cypher~(TM) stent and Bx sonic stent in the same artery.The target vessels were ablated after angiography and observed by scanning electron microscope at one month and three months after implantation of stents respectively.Result At one month,target vessels of Cypher~(TM) stent were covered with endothelium partly and platelet activation was observed.Target vessels of Bx sonic stent were entirely covered with endothelium and platelet activation wasn't observed.Three months later,target vessels of Cypher~(TM) stent and Bx sonic stent were entirely covered with endothelium and there was no platelet activation on the vessel surface for both groups.Conclusion Endothelium regeneration after implantation of Cypher~(TM) stent was much slower than that of implantation of bare metallic stent,and at three months,the endothelialization was finished.Intensive anti-platelet therapy should be considered after implantation of Cypher~(TM) stent.

Objective: This study explored the predictive effects of psychological stress in early and middle puberty on subsequent emotional and behavioral problems. Methods: A cohort of 911 boys and girls from primary and secondary schools in Chongqing was studied. Psychological stress was measured in early and middle adolescence, and the levels of emotional and behavioral problems were determined in one year follow up. Logistic regression analysis was used to analyze the effects of psychological stress in early and middle adolescence on subsequent emotional and behavioral problems. Results: The average psychological stress score in early and middle adolescence was (27.96±24.95), and the average scores of internalized and externalized behavioral problems and overall problems one year later were (8.21±8.46)(6.21±5.87) and (37.90±24.68) respectively. When grouped by gender, these scores, as well as the detection rate of anxiety and depression, withdrawal, physical discomfort, thinking problems, internalization problems and overall problems, were higher in girls than boys (t/Z=-6.38，-5.63，-6.06，-3.74，-6.80，-5.47， χ 2=15.88，12.01，3.92，6.64，24.67，15.88， P <0.05). Logistic regression showed that the psychological stress scale score was a risk factor for internalized and externalized behavioral problems and overall problems( P <0.05). Conclusion Psychological stress levels in early and middle puberty have a positive predictive effect on emotional and behavioral problems in the following year.

Objective: To explore whether psychological stress in early and mid puberty is predictive of adolescent healthrisk behaviors. Methods: In April 2018, a total of 1 046 primary and secondary school students from grade 5 to 8 were recruited through purposive sampling. The basic information and psychological stress of the respondents were collected by questionnaire and physical examination. In addition, self reports of health risk behaviors were collected in October 2018. Logistic multiple regression analysis was used to analyze the relationship between psychological stress and health risk behaviors of adolescents. Results: The reported health risk behaviors of the surveyed adolescents ranking from high to low were unreasonable physical activity (62.05%), food preferences (24.76%), fighting (21.03%), suicidal ideation (17.02%), ideation of running away from home (9.85%), low mood (8.80%), smoking (4.21%), drinking (3.73%), gambling (3.63%) and internet addiction (3.06%). Logistic multiple regression analysis showed that the high level of psychological stress was a risk factor for fighting ( OR =1.02, 95% CI =1.01-1.03), suicidal ideation ( OR =1.02, 95% CI =1.01-1.03), low mood ( OR =1.02, 95% CI =1.01-1.03), ideation of running away from home ( OR =1.02, 95% CI =1.01-1.03), unreasonable physical activity ( OR =1.01, 95% CI =1.00-1.02), smoking ( OR =1.02, 95% CI = 1.00-1.03), internet addiction ( OR =1.02, 95% CI =1.00-1.03), food preferences ( OR =1.01, 95% CI = 1.00-1.02) and gambling ( OR =1.02, 95% CI =1.01-1.04)( P <0.05). Conclusion Unreasonable physical activity and food preferences are most common health risk behaviors among adolescents. Psychological stress during early to middle puberty is predictive of adolescent health risk behaviors.

Objective: To explore the relationship between the emotional and behavioral problems and family relations of firstborn children from an urban area of Chongqing during the role transition period, to provide a scientific basis for creating a good family relationship and to reduce the occurrence of children s emotional behavior problems. Methods: Totally 1 359 mothers of children in the role transition period were recruited from obstetric clinics at two Grade A hospitals in Chongqing, and mothers with informed consent using self designed questionnaire and parental version of the Child Behavior Checklist (CBCL). It was analyzed the association between family relationships and the emotional/behavioral problems of firstborn children of different ages. SPSS statistical software (version 20) was used for the descriptive statistical analysis, chi square test, and multiple linear regression analysis. Results: The parental relationship (56.6%), mother child relationship (80.4%) and father child relationship (64.9%) of most firstborn children was good, and 61.7% of families enjoyed a harmonious atmosphere. For the age groups 1.5-5 years and 6-13 years, the respective CBCL total scores were (18.22±13.63) and (24.20±17.52), and the detection rates were 10.7% and 10.5%, respectively. The results of the multiple linear regression analysis showed that, for both age groups, firstborn children who had good parental relationships, good mother child relationships, good father child relationships, and a harmonious family atmosphere exhibited fewer internalizing, externalizing and overall emotional behavior problems ( P <0.05). Conclusion The total detection rate of emotional and behavioral problems among firstborn children in the role transition period, which is low. Good parental relationships, good parent child relationships, and a harmonious family atmosphere are protective factors against emotional and behavioral disorders in firstborn children during the role transition period.

Objective: To compare the emotional and behavioral characteristics of preschool firstborn children across the transition to siblinghood with the only children in a district of Chongqing. Methods: Data were derived from a cohort study on firstborn children transition to siblinghood. Totally 882 firstborn children (3.58±1.25) years old and 329 only children (3.98±1.02) years old were recruited from two hospitals and 19 kindergartens in a district of Chongqing, respectively. Their mothers filled out the parental version of Child Behavior Checklist (CBCL) and self designed questionnaire. Results: The overall scores of emotional and behavioral problems of firstborn children and only children were (25.17±18.82) and (31.75±18.64). The results of rank sum test showed that the scores of the firstborn children in all dimensions, internalizing and externalizing behavior problems and overall emotional and behavioral problems were lower than those of the only children ( P <0.05). Chi square test results showed that the detection rates of internalizing, externalizing and overall emotional behavior problems in only children(12.77%, 13.07%, 14.59%) were higher than those in firstborn children(8.28%, 8.28%,8.16%) ( χ 2=5.62, 6.36, 11.09, P <0.05). Multiple linear regression showed that after adjusting for age, gender, family economic pressure, temperament type of children, family atmosphere and family type, the scores of internalizing problems, externalizing problems and overall problems of the only children were still higher than those of the firstborn children ( B =1.54, 2.32, 6.81, P <0.05). Conclusion The firstborn children across the transition to siblinghood show less emotional and behavioral problems than the only children, and the scores and detection rates of all types of emotional and behavioral problems of the firstborn children are lower than those of the only children.

Objective: To explore the influence of parental company before and after having second child on the emotional and behavioral problems of preschool firstborn children, and to provide a reference for emotional problems prevention of firstborn children. Methods: Questionnaire survey was conducted among mothers of firstborn children who met the inclusion and exclusion criteria from two Grade A hospitals in Chongqing from March to December 2019. The average age of 845 firstborn children was (3.56±1.24) years old. The chi square test was used to compare the general characteristics and parent company of boys and girls. Rank sum test was used to compare the differences of emotional and behavioral problems of firstborn children with different parents company. Multiple linear regression models were used to compare the differences of internalizing, externalizing and total problems in firstborn children with different parents company. Results: After having the second child, parents spent less time with their firstborn children, among which the decrease of the accompanying time of the mother and the total accompanying time of the parents were statistically significant( Z =3.76, 2.86, P <0.05). The scores of internalizing problem were higher when the mother s company time decreased ( B=1.18, 95%CI = 0.01 -2.36) and the parents total company time decreased( B=1.41, 95%CI =0.33-2.48). The scores of the externalizing problem ( B=1.25, 95%CI =0.10-2.39) and the total problem ( B=4.31, 95%CI =1.05-7.57) with the decrease of parents total company time were higher. Conclusion Parent company has an important influence on the emotional and behavioral problems of firstborn children. Firstborn children whose parents accompanying time decreases after having second children scored higher on emotional and behavioral problems.

Objective: To explore the status of depressive symptoms among adolescents in different pubertal stages in a district of Chongqing. Methods: A self designed questionnaire and Children s Depression Inventory were administered among 1 001 students in a district of Chongqing. Score and associated factors of depressive symptoms of adolescents through descriptive statistical analysis, Chi square test, analysis of variance and Logistic regression analysis. Results: The detection rate of depressive symptoms among adolescents in a district of Chongqing was 17.3%, and the total score of depression scale was (12.02±6.72). There were statistically significant differences in the inefficiency subscale score of boys across stages of genital, pubic hair, armpit hair and beard, and whether the first spermatorrhea has occurred( t/F =7.08，5.46，5.18，4.21，5.84， P <0.05), while significant differences were found in the anhedonia and inefficiency subscales scores among girls across different stages of breast, pubic hair and armpit hair, and whether menarche has occurred( t/F =19.43，4.92，3.98，7.35， P <0.05). Multivariate Logistic regression analysis showed that the absence of first spermatorrhea and menarche were associated with lower prevalence of depressive symptoms in boys and girls ( OR=0.60, 95%CI=0.36-0.98; OR=0.46, 95%CI =0.22-1.00), while pubic hair development was associated with higher prevalence of depressive symptoms in girls ( OR=9.58, 95%CI =1.28- 71.71 ). Conclusion The detection rate of depressive symptoms among adolescents is relatively low. Boys who have had the first spermatorrhea, and girls with advanced pubic hair development or have had menarche are more likely to suffer from depressive symptoms.

OBJECTIVE: To investigate the protective effect of quercetin against lipopolysaccharide (LPS)- induced acute kidney injury (AKI) in mice and explore its mechanism. METHODS: Forty male BALB/c mice were randomly divided into control group (with saline treatment), 15 mg/kg LPS group, and quercetin-treated groups with intragastric quercetin treatment (once daily for 3 consecutive days) at low (25 mg/kg) and high (50 mg/kg) dose prior to 15 mg/kg LPS injection. LPS was administered by intraperitoneally injection 1 after the last gavage of quercetin. The mice were sacrificed 24 h after LPS injection for analysis of kidney pathologies, blood urea nitrogen (BUN) and creatinine levels; serum levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6 were detected by ELISA, and the expressions of Toll-like receptor-4 (TLR4), MyD88, TRAF-6 and NF-κBp65 in the kidney were detected by Western blotting. RESULTS: Quercetin significantly lessened renal pathologies, lowered BUN and creatinine levels ( < 0.05) and inhibited TNF-α, IL-1β, and IL-6 production in mice with LPS-induced AKI ( < 0.05). Pretreatment with quercetin also significantly inhibited TLR4, MyD88, and TRAF-6 expressions and NF-κBp65 activation in the kidneys of the rats with LPS challenge ( < 0.05). CONCLUSIONS Quercetin pretreatment can protect mice against LPSinduced AKI by inhibiting TLR4/NF-κB signaling pathway.
Acute Kidney Injury ; Animals ; Antioxidants ; pharmacology ; Lipopolysaccharides ; Male ; Mice ; Mice, Inbred BALB C ; NF-kappa B ; Quercetin ; pharmacology ; Rats ; Toll-Like Receptor 4 ; drug effects ; Tumor Necrosis Factor-alpha ; drug effects