Objective : To explore the mechanism of hippocampal corticotropin-releasing hormone ( CRH) receptor type 1 ( CRHR1 ) in chronic stress-induced learning and memory impairment in early aged mice． Methods: C57BL /6J mice aged 12 －14 months were divided into two groups according to gender，and then divided into wild type (WT) group and hippocampal CRHR1 conditional gene knockout (KN) group according to genotype．Mice in each group were randomly divided into control group and stress group，and the stress group was subjected to chronic unpredictable stress ( CUS ) for 30 days． Genotyping of mice was performed using polymerase chain reaction ( PCR) ，agarose gel electrophoresis and real-time fluorescence quantitative PCR (RT-qPCR) ．The new object rec- ognition experiment and Morris Water maze measured learning and memory ability．Golgi-Cox staining was used to observe damage to hippocampal neuronal dendrites． The protein expressions of target protein of rapamycin (mTOR) ，p-mTOR (Ser2448) ，ribosomal protein S6 kinase ( p70S6K) and p-p70S6K ( Thr389 / Thr412 ) were detected by Western blot．Serum levels of corticotropin releasing hormone ( CRH) were measured by ELISA. Results : Compared to mice without chronic stress，the cognitive coefficient of WT stress groups decreased after chron- ic stress，and the difference was statistically significant (P ＜0. 05) ，while there was no significant difference in cognitive coefficient of KN stress groups before and after chronic stress．Compared with the WT stress group，the escape latency of the WT control group was shortened (P＜0. 05) ，and the number of crossing the platform and tar- get quadrant increased (P ＜0. 01) ，and there was no significant difference in the KN groups above． Compared with the WT control group，the WT stress group had a significant reduction in the neuronal complexity in the hipp- ocampal CA1，CA3 and DG regions (P ＜0. 05) and significant reductions in the expression of p-mTOR and p- p70S6K in the hippocampus (P＜0. 05) ．There was no significant difference in the expression of p-mTOR between the KN stress group and the KN control group (P＞0. 05) ，except that the expression of p-mTOR in the hippocam- pus of the female group decreased (P＜0. 05) ．In addition，the serum level of CRH in the stress group was higher than that in the control group (P＜0. 01) ． Conclusion Hippocampal CRHR1 regulates learning and memory im- pairment and neuronal dendrite damage in early aged mice induced by chronic stress．The mechanism may be that high levels of CRH induced by chronic stress cannot bind to CRHR1 receptor，thereby enhancing the expression of down-regulated mTOR / p70S6K signaling pathway.

Objective : To investigate the sex difference of the effects of chronic pregnancy stress on depression-like behavior in offspring adolescent mice and whether the amygdala is involved in mediating depression-like behavior and its possible mechanism． Methods : Male and female of C57BL /6J mice were put in cage together．Pregnant mice were randomly divided into normal control group ( CON group) and chronic pregnancy stress group ( CPS group) ．The day of delivery was recorded as post-natal day(PND0) ．The offspring of different groups were divided into Female group and Male groupaccording to sex，respectively．From PND35，the depressive-like behavior of off- spring was monitored in different groups．Morphological structure of basolateral amygdala (BLA) cone neurone was observed by Golgi-Cox staining，and apoptosis of BLA neurone was detected by TUNEL．Serum corticotrophin-relea- sing hormone ( CRH) was detected by ELISA．The level of protein associated with amygdala mammalian target of rapamycin (mTOR) and phosphorylated mammalian target of rapamycin ［p-mTOR ( Ser2448) ］was detected by Western blot. Results : Depression-like behavior was appeared in different sexual offspring by chronic pregnancy stress，and there was an interaction between chronic pregnancy stress and gender．In the forced swimming test，the immobility time of offspring in the CPS group prominently increased(Female: P＜0. 05，Male: P＜0. 001) ．Interest- ingly，compared with female offspring ，despairing behavior of male offspring was much more clearly observed in CPS group(P＜0. 05) ．Compared with offspring of CON group，the rate of sucrose preference was significantly re- duced in the female offspring of CPS group(P＜0. 05) ，while no obvious difference was observed in the male off- spring．Compared with the CON group，the density of neuronal dendrite branches in the BLA of offspring mice in the CPS group decreased(Female: P＜0. 01，Male : P＜0. 01) and the degree of neuronal apoptosis increased( Fe- male: P＜0. 001，Male : P ＜0. 001) ，the expression level of p-mTOR in amygdala of offspring mice in CPS group significantly decreased(Female: P＜0. 001，Male: P＜0. 001) ．Chronic pregnancy stress increased the serum CRH level of offspring mice(P＜0. 001) ，and the gender had significant influence on serum CRH level，the serum CRH level of female in CPS group was higher than that of male(P＜0. 05) ． Conclusion Chronic pregnancy stress leads to depression-like behavior in offspring adolescent mice，and the depression-like behavior has gender differences. In addition，chronic pregnancy stress leads to dendrite atrophy and apoptosis of BLA neurons in offspring mice，and the mechanism may be that the activation of mTOR in the amygdala of offspring mice is inhibited．CRH may be in- volved in mediating sex differences in depression-like behavior and BLA neuron damage in offspring induced by chronic pregnancy stress.

OBJECTIVETo analyze the associations between pregnancy-related anxiety and the prevalence of subthreshold autism trait (SAT) in preschool children.

METHODSBaseline data came from the Ma'anshan Birth Cohort Study, a part of the China-Anhui Birth Cohort Study (C-ABCS). All the participants were enrolled among pregnant women who received prenatal health care in 4 municipal medical centers during Oct. 2008 to Oct. 2010. A total of 5 084 pregnant women were recruited at the beginning and 4 669 singleton live births were included until childbirth. The situation about pregnancy-specific anxiety during trimester and third trimester of women were evaluated by Pregnancy-specific Anxiety Questionnaire (PAQ). Between April 2014 and April 2015, the cohort was followed up again, and the Clancy Autism Behavior Scale (CABRS) filled out by parents was used for telling the SAT children from the healthy children among 3 663 preschool children. Univariate and binary regression model was used to estimate associations between the pregnancy-related anxiety during trimester and third trimester and the subthreshold autism trait in children.

RESULTSDuring the pregnancy, the detected rates of women with pregnancy-specific anxiety in trimester and the third trimester were 25.5%(935/3 663), 13.9%(501/3 592) respectively, and the detected rate of maternal pregnancy-specific anxiety in both periods was 7.7%(278/3 592). There were 290 positive children with SAT and the detection rate was 7.9%. After controlling possible confounding factors including children's genders, place of residence, supplement folic acid during pregnancy, preterm birth, exposure to second-hand smoke during pregnancy, the father (mother) cultural levels, the father (mother) nature of work and family income, the results of multinomial logistic regression analysis showed that maternal pregnancy-specific anxiety in trimester was the risk factor for SAT in preschool children (OR=1.51, 95% CI: 1.11-2.04), and there was no association between maternal pregnancy-specific anxiety in the third trimester and SAT in preschool children (OR=1.36, 95% CI: 0.82-2.22). Compared with the single function of maternal pregnancy-specific anxiety in trimester or the third trimester for SAT in preschool children, maternal pregnancy-specific anxiety in both periods presented a joint action that increasing the risk for SAT (OR=2.02, 95% CI: 1.36-2.98).

CONCLUSIONMaternal pregnancy-related anxiety was a risk factor for subthreshold autism trait in preschooler children. Pregnant women should try to keep a good mental state to create a good environment for fetal growth.

Anxiety ; epidemiology ; Autistic Disorder ; epidemiology ; Child ; Child, Preschool ; China ; Cohort Studies ; Dietary Supplements ; Female ; Humans ; Infant, Newborn ; Pregnancy ; Pregnancy Complications ; psychology ; Pregnancy Trimester, Third ; psychology

Objective To investigate the correlation between body mass index (BMI),waist circumference (WC)and sperm quality parameters of male infertility.Methods 303 cases of male infertility patients including outpatients and inpatients were selected to measure height,weight and WC,and to group according to BMI standards,and sperm quality parameters of each group were detected simultaneously.Results Overweight,obese and severely obese group sperm concentration had very significant difference compared with normal weight group (t = 3.941,3.782,5.632,P <0.01);Overweight,obese and se-verely obese group total sperm count had significant difference compared with normal weight group (t = 2.117,2.655, 8.591,P <0.05 or 0.01).No difference between the normal weight and low weight group (t=0.668,P >0.05),sperm con-centration and total sperm count decreased with increasing BMI.Low weight group,overweight,obese and severely obese group were higher than normal sperm immobile body recombination,there were significant differences (t = 2.847,8.592, 8.472,5.380,P <0.05).Normal sperm morphology rate of abnormal weight and normal weight groups were significantly lower comparison,the difference was significant (t=2.356,5.968,5.156,5.993,P <0.05).Abnormal body weight of each group of sperm motility and normal weight group were significantly lower comparison,and there were significant differences (t=2.847,8.593,8.472,5.380,P < 0.05).Forward movement rate abnormal and normal body weight of each group re-structuring comparison were significantly lower,and there was a significant difference (t=2.432,11.816,9.588,13.528,P<0.05).Overweight,obese and severely obese group of sperm DNA fragmentation index were higher than the normal weight group,which the data showed a significant differences (t=6.168,7.238,9.309,P <0.05),while there was no statis-tical significance between low weight group and normal weight group (P >0.05).Conclusion Studies showed that the ab-normality of BMI and obesity could cause the changes in sperm parameters which might be an important mechanism of male infertility.

Objective To explore whether prenatal stress promotes formation of chronic stress-induced hippocampal amyloid β (Aβ) protein in 6-month-old male offspring mice and its mechanism.Methods The APPswe/PSIdE9 double transgenic mice were divided into 4 groups according to the prenatal stress and offspring mice stress:prenatal control-offspring control group (CC group),prenatal control-chronic offspring stress group (CT group),chronic prenatal stress-offspring control group (TC group),and chronic prenatal stress-chronic offspring stress group (TT group) (n=18).The number of amyloid plaques in brains was checked using Congo red staining.ELISA was used to examine the hippocampus levels ofamyloid-β proteins (Aβ1-42 and Aβ1-40) in the offspring mice;β-site APP-cleaving enzyme 1 (BACE1) activity was detected using fluorospectrophotometry.Additionally,Western blotting were used to observe the expression levels of phosphorylated eukaryotic initiation factor 2α (p-eIF2α),phosphorylated protein kinase R [PKR]-like ER kinase (p-PERK),glucose-regulated protein 78 (Grp78) and β-site BACE1 in the hippocampus.Results As compared with that in the CC group,the number of amyloid plaques in brain in CT,TT and TC groups was increased.The expressions of p-eIF2α,p-PERK,Grp78,BACE1,Aβ1-40 and Aβ1-42 in the hippocampus of CT group were significantly increased as compared with those in the CC group (P＜0.05).The expressions of p-eIF2α,p-PERK,Grp78,BACE1,Aβ1-40 and Aβ1-42 in the hippocampus of TT group were further significantly increased as compared with those in the CT group (P＜0.05).There was no significant difference in BACE1 activity among the different groups (P＞0.05).Conclusion The prenatal stress can promote the formation of hippocampal Aβ protein induced by chronic stress in 6-month-old male offspring mice,whose mechanism may be that prenatal stress aggravates hippocampal neurons endoplasmic reticulum stress,activates the PERK,then causes eIF2 alpha phosphorylation,and finally promotes BACE1 expression.

Objective To determine whether chronic stress could potentiate learning and memory impairment in old mice, and, if so, what the underlying mechanism is. Methods Sixty male mice were divided randomly into control group and chronic stress group. Mice in stress group were stressed everyday by one of the stressors including cold exposure, restraint, level shake and so on. The ability of learning and memory was determined by Morris water maze test, and the histopathologic changes in CA3 field of the hippocampus were examined under a light micro-scope. Serum corticosterone level was determined by enzyme-linked immunosorbent assay. Western blot was per-formed to determine the expression of β-site amyloid precursor protein-cleaving enzyme 1 and Aβ1-42 in hippocam-pus of the brain. Results Compared with the control group, the results showed that chronic stress could increase the escape latency and swimming distance of old mice during training session in the Morris water maze test. The neuropathological changes were characterized by the decreased neuron number,soma shrinkage and condensation,or nuclear pyknosis in the CA3 field of hippocampus in the stress group. On the other hand, the expression of Aβ1-42 and BACE1 protein in hippocampus were increased, as well as the serum corticosterone concentration in the stress group. Conclusion Chronic stress can potentiate learning and memory impairment and pathological damage in CA3 field of the hippocampus in old mice, which may be related to chronic stress up-regulated the levels of BACE1 and Aβ1-42 mediated by corticosterone.

Objective: To study the effect of total flavonoids of astragalus (TFA) on adjuvant arthritic (AA) rats and its mechanism. Methods: The volume of non-injected hind paw of AA rats, serum malondialdehyde (MDA) content, interleukin-1(IL-1) and nitrite (NO - 2) produced from articular synoviocytes were measured. Results: It was obseved that serum levels of MDA and the levels of IL-1 and NO - 2 from synoviocytes increased in AA rats, and the degree of the secondary inflammatory reaction of AA rats appeared to be directly correlated with serum levels of MDA and IL-1. Treatment of whole (d0～27) or partial (d12-18 or d18-24) course of AA rats with TFA (20 mg/kg/d,ig) could not only markedly inhibit the inflammatory reaction in AA rats, but also reduce their enhanced serum lipid peroxides (LPO), IL-1 and NO production from synoviocytes. Conclusion: TFA has significant therapeutic effects on AA rats, which might be related to both of anti-oxidative effect and the reduced production of IL-1 and NO from synoviocytes.

Aim To study the effect of EA on the injury induced by ?-amyloid protein(A?) in primary cultures of rat hippocampal neurons. Methods The protective effect of EA on A?_25-35 induced neurons injury was observed by LDH release rate, MTT, LSCM and TUNEL. Results A?_25-35 could induced cell death in rat primary hippocampal neurons. Four hours pretreatment with 20 mg?L-1, 40 mg?L-1 EA exerted the protective effect on rat primary hippocampal neurons from A?_25-35 induced injury. Conclusion EA had protective effects against injury induced by A?_25-35 in rat primary hippocampal neurons to some certain,which probably related with decreasing the level of calcium.